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Open Journal of Genetics, 2011, 1, 38-47 OJGen

doi:10.4236/ojgen.2011.13008 Published Online December 2011 (http://www.SciRP.org/journal/ojgen/).

Effect of temperature on crossing over in the mus309 mutant,
deficient in DNA double-strand break repair, of Drosophila
melanogaster suggests a mechanism for crossover interference
Petter Portin
Laboratory of Genetics, Department of Biology, University of Turku, Turku, Finland.
Email: petter.portin@utu.fi

Received 16 September 2011; revised 27 October 2011; accepted 19 November 2011.

ABSTRACT lication of the chromosomes. Meiosis involves pairing of
the homologous chromosomes and, exchange by cross-
Crossing-over frequencies, crossover interference, ing over of genetic material between them resulting in
recombination frequencies and map distances were the formation of chiasmata. A chiasma is a sufficient
compared in the cv-v-f region of the X chromo- condition for the segregation of homologous chromo-
some of Drosophila melanogaster in females bearing somes, which leads to the reduction of the chromosome
either wild type 3rd chromosomes (control) or hav- number from diploid to haploid. The alternation of mei-
ing the DNA double-strand break repair deficient osis and fertilization leads to the recombination of ge-
mus309D2/mus309D3 mutant constitution in the 3rd netic material.
chromosomes (experiment), and raised in three dif- An important phenomenon associated with crossing
ferent temperatures viz. 18˚C, 25˚C and 29˚C. In over is crossover interference, i.e. the fact that multiple
addition, the fecundity of the females was also crossovers are less frequent than would be expected on
measured. In the control crosses none of the mean the basis of random coincidence of single crossovers
values of the parameters measured was dependent [1-3]. Crossover interference has recently garnered con-
on the temperature, whereas in the experimental siderable attention because it may be responsible for the
crosses all the parameters, except for the frequency occurrence crossovers called obligate crossovers, and
of true single crossovers in the cv-v interval, the thus for the formation of obligate chiasmata.
recombination frequency of the v and f markers, The term “obligate crossover” refers to the fact that, in
and the coefficient of coincidence, changed due to most species, it is rare to find chromosomes that do not
the effect of temperature. When comparing the ge- undergo crossing over. For example, in Drosophila, there
notypes studied, a significant difference between is usually one chiasma per chromosome arm. The feature
them was observed in all the parameters measured, of the obligate chiasma is biologically sensible because
apart from the frequency of the true single cross- it ensures the disjunction of homologous chromosomes.
overs in the cv-v interval. These results support the
counting number model of the mechanism of inter- 1.2. Models of Crossover Interference and the
ference based on the genetic distance, but are in Aim of the Present Study
contradiction with the models based on physical Models of crossover interference can, in principle, be
distance. divided into two different categories. The first category
of models, called genetic models [4], assumes that inter-
Keywords: Chiasma; Chromosome; Map Length; Meio- ference is dependent on the genetic (i.e. linkage map)
sis distance (Morgans) between adjacent crossovers. To my
knowledge, currently only one model, called the ‘count-
1. INTRODUCTION ing model’, falls into this category [4,5].
1.1. General Introduction The second category of models, which may be called
physical models, hypothesize that crossover interference
Meiosis is a fundamental process in all sexually repro- is dependent on the physical distance (microns or base
ducing eukaryotes, consisting of two successive divi- pairs) between the adjacent crossovers. In general, these
sions of the meiocyte nucleus but only one cycle of rep- models, which are many, suggest that some kind of

Published Online November 2011 in SciRes. http://www.scirp.org/journal/OJGen

manner similar to its orthologues in other organisms. and the coefficient of co- incidence. These results support the counting ion (v. was that following the while the DSBR pathway remains functional [25]. More specifically. or D-loop. Crossing. the fecundity of the females was also mea.22]. were made at three different temperatures. Molecular Models of Crossing Over and the homozygous cv v f females were crossed with + + +/Y mus309 Gene males. to non-crossover products and the latter to both cross- mosomes (control) or having the DNA doublestrand over and non-crossover products [16.7). there Subsequent DNA synthesis and second end capture form is a stop codon between the sequence motifs encoding the Copyright © 2011 SciRes. changed due to the effect of temperature. P. crossovers [14. crossovers in the cv-v interval. a RecQ helicase [18-21] sured. (DSBR) pathway.13. in co-operation with phenotype. Both the mus309 alleles used carry mutational changes end-invasion intermediate (SEI). The occurrence of DSBs is fol. 2. In the experimental crosses. 1 . the mus309 locus on the stitution in the 3rd chromosomes (experiment). which is then resolved to form either crossovers or non- The aim of the present study was to distinguish be. none of the mutants more DSBs are repaired as crossovers by the mean values of the parameters measured was dependent DSBR pathway. in a in three different temperatures viz. by doing so one can tell some. the oocyte between the two alternative pathways of DSB thing about the mechanism of crossing over and cross. arguments of several earlier authors [6-8]. whereas in the experimental crosses ratio can be expected.24]. but are in contradiction with the cated below. In the control fly crosses. it is known that the product of the mus309 mutation on crossing over and crossover interference locus is involved in the SDSA pathway of the repair of was chosen as the experimental design in the first place the DSBs [23. mus309D2/mus309D3 Present molecular models of meiotic crossing over and females were crossed with cv v f/Y males. it is also known because temperature is easy to control. In mus309D2. more important reason. viz.1.56. a physical that could potentially impair or abolish at least the heli- structure called a displacement loop. 1 . vermil- the cv-v interval.33. Following this. over interference in particular. 18˚C. crossover interference.15]. which in Drosophila is en. The combined effect of temperature and the mus309 In particular. if in mus309 It was observed that in the control crosses. 1 . a significant difference The effect of temperature on crossing over frequency between them was observed in all the parameters studied. In Drosophila melanogaster. +/+ models based on physical distance. 25˚C and 29˚C. repair. The former pathway leads exclusively nogaster in females bearing either wild type 3rd chro. The same is also true for the mus309 orthologue. 18˚C  lowed by formation of heteroduplex DNA and rejoining 1˚C. liminary cross: cv v f. and that in mus309 mutants the SDSA pathway is blocked. Consequently. ca. mammalian BLM locus included. a change in the crossover/non-crossover on the temperature. and as ex. Portin / Open Journal of Genetics 1 (2011) 38-47 39 physical signal travels along the bivalent and determines a structure known as a double Holliday junction (dHJ). in yeast [26]. and. since fewer non-crossovers are all the parameters except for the frequency of true single produced. the Sgs1 locus. 25˚C  1˚C and 29˚C  0. the mus309 gene seems to control the choice made by plained in Discussion. Tb females cross- talyzed most likely in all eukaryotes by the topoisom. cv v f/+ + +. The second. Both the preliminary and the actual crosses other enzymes [10-13].6) and identified on the basis of their non-Tubby coded by the mei-W68 gene [9]. . break repair deficient mus309D2/mus309D3 mutant con. MATERIALS AND METHODS quency of the v and f markers.17]. Two alternative pathways for the repair of the DSBs over frequencies.3.17. however. were crossed with their cv v f/Y brothers. ed with + + +/Y.5˚C.0) and forked (f. OJGen . Tb males (Tb. cv v f/+ + +. mus309D3/TM6.90. Thus. females derived from the preliminary cross in which 1. accordingly. mus309D2/TM6. Experimental Procedures comparing the genotypes studied. the recombination fre. Tubby 3 erase-like Spo11 protein. When 2. case function of the MUS309 protein.7) markers was number model of the mechanism of interference based compared in the experimental and control crosses indi- on the genetic distance. recombination are known: the synthesis-dependent strand annealing frequencies and map distances were compared in the (SDSA) pathway and the double-strand-break repair cv-v-f region of the X chromosome of Drosophila mela. is formed. and crossover interference in the X chromosome in the except for the frequency of the true single crossovers in regions between the crossveinless (cv. The experi- gene conversion suggest that crossing over is initiated by mental females were derived from the following pre- the formation of DNA double-strand breaks (DSBs). the In addition. the distribution of crossovers. is involved in DSB repair [16. of the ends created in the breakage involving a single. and raised right arm of chromosome three (86F4) encodes. tween the explanatory values of these models.

which is a maxi- Because of the semi-sterility of the females. Student’s t-test and the binomial t-test were Single Crossovers employed. This is also a maximum were as follows: In 18˚C 19 bottles. 3. the three classes of double-exchange tetrads. syrup. in 25˚C 19 bottles. i. the ex. in addition to another 2. The progeny were raised on a stan. 2. The results of the control crosses. and v and f.2. The parental and n is the total number of flies. Measurement of Interference amino acid substitution close to the C terminus [27]. cording to the following formula [31]. OJGen . The num. prog- eny was not derived from every culture bottle. respectively. the true frequency of single crossovers. females were allowed to lay eggs in the culture bottles for The variance of C was calculated according to the four days and were transferred thereafter together with the following formula [31] males into fresh culture bottles for two more days of c  1  ca  cb  cab  2c 2 ab  egg-laying. +/+ females crossed with cv v f/Y. C. 2-. has a glutamic acid to lysine substitution in over classes. was calculated ac- mus309D3 is semi-sterile [27-29]. double crossovers from that of each of the single cross- for its part.  w  x  w  y  trol crosses were single-female cultures. and in 29˚C 20 bottles. Phenotype of the Total number Number of progeny progeny of flies +++ cv v f cv + + +vf cv v + ++f cv + f +v+ Temperature 18˚C 607 524 144 187 179 181 23 23 1868 25˚C 680 549 143 206 125 219 21 22 1965 19˚C 618 429 163 168 138 164 27 31 1738 ∑ 1905 1502 450 561 442 564 71 76 5571 Copyright © 2011 SciRes. occur in a 1:2:1 ratio [30]. and in 29˚C 18 bottles. and the respec- calculated by subtracting the observed frequency of ve distribution in the experimental crosses in (Table 2). (1) 3 females were mated with 3 . Thus.e. In the statistical analyses of the results. Because of accidental death of the females. distribution of progeny raised at three different temperatures (18˚C. crossovers. Portin / Open Journal of Genetics 1 (2011) 38-47 third and fourth helicase motif of the protein. 3.4. agar-agar and both dried and fresh yeast.and 4-strand doubles. +/+ males. x and y are the numbers of flies which were dard Drosophila medium consisting of semolina. interference. progeny was the result of a total of six V (cˆ)   . (2) n  ab  days. Calculation of the Frequency of the True of variance.40 P. and v and f. respectively. likelihood equation.3. whereas the con. 25˚C and 29˚C) of cv v f/+ + +. mus309D3. where a and b are the recombination frequencies of cv bers of bottles which gave progeny in the control crosses and v. Assuming no chromatid are given in the appropriate table (Table 3). the conserved helicase II motif. was in the control crosses is given in (Table 1). one-way analysis 2. single crossovers for cv and v. one in each interval. The distribution of the progeny into different phenotypic the number of single crossovers that resulted from classes in each temperature as well as their total numbers meioses with only one exchange in the cv-v-f region. Statistical Methods in 25˚C 20 bottles. Table 1. RESULTS Therefore. Similarly. It has been demonstrated that the genotype mus309D2/ The coefficient of coincidence. The same number (20) of crosses was made in both the control and the where w is the number of flies which were double experimental series. the results of the binomial t-tests exchanges. The results of the analyses of variance as well as those of the student’s t-test including their signify- Some of the observed single crossovers in the cv – v and cances are always given in the figure legend of the rele- v – f intervals actually result from meioses that have two vant figure.5 males. mum likelihood equation perimental crosses were carried out in cultures in which wn cˆ  . The respective numbers in the experimental crosses were as follows: In 18˚C 19 bottles.

The significance of the difference of the tioned was calculated according to the Student’s t-test. The mean frequency together with its standard deviation given and indicated by the red diamond (♦). The effect of temperature on the frequency of the Figure 1. genotype D2/D3: F = 34. the P-values for the test of heterogeneity = 114 and P = 0. df2 = 56.0001). It appears also that the mean mental females (P = 0.19. the fecundities The frequencies of the true single crossovers in the of the females. the result of which was as follows: t = 0. In addition. the mean values mentioned was calculated according to the Stu- result of which was as follows: t = 14. and that for the test of the difference be- types in the upper right corner of the figure.79. The effect of temperature on the fecundity of the true single crossovers between the cv and v markers of the X mus309+/mus309+ (+/+) and mus309D2/mus309D3 (D2/D3) chromosome in the progeny of the mus309+/mus309+ (+/+) females kept in three different temperatures. As appears perimental females are given in (Figure 2). viz.0001. in each temperature for the control and cv-v interval in each temperature for the control and ex- experimental crosses is given in (Figure 1). the fecundity of the control females was was temperature sensitive in neither group of females. df2 = 53. P < 0. and that for the test of the difference between the geno. 18˚C.83. OJGen . the P-values for the test of are given below the group of columns for the respective geno. +/+ males. The frequency from the figure. df = 115 and P < dent’s t-test.85. Copyright © 2011 SciRes. distribution of progeny raised at three different temperatures (18˚C. Portin / Open Journal of Genetics 1 (2011) 38-47 41 Table 2. 0. per culture bottle and the vertical lines their standard devia- gether with its standard deviation for both genotypes is also tions.70. P = 0. Phenotype of Total number Number of progeny the progeny of flies +++ cv v f cv + + +vf cv v + ++f cv + f +v+ Temperature 18˚C 440 387 143 168 128 163 34 37 1500 25˚C 655 540 189 206 216 261 31 41 2139 19˚C 888 859 245 280 279 313 51 36 2951 ∑ 1983 1786 577 654 623 737 116 114 6590 The number of progeny per female. mus309D2/mus309D3 females crossed with cv v f/Y. 25˚C and mus309D2/mus309D3 (D2/D3) females raised at three dif- and 29˚C. The mean number to.36. df1 = 2. P. df1 = 2. heterogeneity are given below the group of columns for the type.Testing of the heterogeneity of the results inside each carried out using the one-way analysis of variance. i.0987. respective genotype. Testing of the for both genotypes is also given and indicated by the red dia- heterogeneity of the results inside each mus309 genotype was mond (♦). The results were as follows: genotype +/+: F = 0. The results mus309 genotype was carried out using the one-way analysis were as follows: genotype +/+: F = 0. 18˚C. In addition.24. P = of variance.0001. tween the genotypes in the upper right corner of the figure.9215). genotype D2/D3: F = 0. whereas that of the was there any difference between the control and experi- experimental females was. Nor not dependent on the temperature. = 2. df1 = 2. df2 = 56. value of the fecundity of the experimental females was significantly lower than that of the control females (P < 0. viz. The results of the experimental crosses. df2 = 53. P = 0. The columns per female in each temperature in the two genotypes and the represent the mean frequencies of crossovers in percentages vertical lines their standard deviations.e. Columns represent the mean numbers of progenies ferent temperatures. df 0. Figure 2. 25˚C and 29˚C) of cv v f/+ + +.0. df1 The significance of the difference of the mean values men.9215. 25˚C and 29˚C.

48 17.032. P < 0.54 24.0039. P = 0. df = 12159. df = 12159.97. df = 115 and addition. +/+ males. region of the X chromosome in each temperature for the perimental crosses are given in (Figure 3). whereas perimental crosses was. the frequency in the control crosses was not As can be seen in the figure.0438 0.0039). Copyright © 2011 SciRes.23 t = 2.54 22.060.0001 Figure 3. df1 = 2. mus309D2/mus309D3 (experimental) females crossed with cv v f/Y. 25˚C and 29˚C.64 ± 0. df = 12159.79 ± 0. 18˚C. df = 114 and P = 0.0362 t = 4.21 3. 25˚C and 29˚C.70 ± 0. The columns represent the the mean frequencies of crossovers in percentages per culture mean frequencies of crossovers in percentages per culture bot- bottle and the vertical lines their standard deviations. df = 12159.0001 Frequency of true single crossovers in the cv – v interval 15. The effect of temperature on the frequency of the true Figure 4. P = 0.42 P. P = 0. Portin / Open Journal of Genetics 1 (2011) 38-47 The frequencies of the true single crossovers in the v The frequencies of the double crossovers in the cv-v-f -f interval in each temperature for the control and ex. and ity are given below the group of columns for the respective that for the test of the difference between the genotypes in the genotype.032. 18˚C.53 t = 4. P = 0.49.70.00. The effect of temperature on the frequency of the single crossovers between the v and f markers of the X chro. df1 = 2. the P-values for the test of heterogeneity are given P = 0.49. genotype D2/D3: F = 3.6134 ± 0. df = 12159. P < 0.04.47 ± 0.85. = 2. The results were as follows: genotype +/+: F = 2.44 t = 0.0023 Frequency of double crossovers in the cv – v – f region 2. df1 = 2. viz. +/+ (control) and cv v f/+ + +.6214 Frequency of true single crossovers in the v – f interval 15.19 ± 0.51 t = 1. It also appears that value of the frequency in question was significantly the mean value of the frequency in question was signifi- higher in the experimental than in the control crosses (P cantly higher in the experimental than in the control = 0. In t-test.51 ± 0.17 ± 0. df2 = 56.30 ± 0. df = 115. double crossovers in the cv-v-f region of the X chromosome in mosome in the progeny of the mus309+/mus309+ (+/+) and the progeny of the mus309+/mus309+ (+/+) and mus309D2/ mus309D2/mus309D3 (D2/D3) females raised in three different mus309D3 (D2/D3) females raised in three different tempera- temperatures.48 ± 31.23 ± 4. the figure. Comparison of different parameters calculated on the basis of the total number of progeny of cv v f/+ + +. and raised at three different temperatures viz.14. P < 0. the P-values for the test of heterogene- below the group of columns for the respective genotype.13 ± 0. and that for the test of the difference between the upper right corner of the figure. df2 = 53.34. OJGen . The mean tle and the vertical lines their standard deviations. It also appears that the mean that of the experimental crosses was.0001 Coefficient of coincidence 0. crosses (P = 0. The significance of the difference of the mean was calculated according to the Student’s t-test.52. crosses was not dependent on the temperature.48 ± 0. The genotype was carried out using the one-way analysis of vari- results were as follows: genotype +/+: F = 2.66 46. In addition. 18˚C.0001 Map distance of the cv and f markers 41. df = 12159.6525 ± 0.49 15. P = 0.0643 Recombination frequency of the v and f markers 20. The columns represent tures. Parameter measured Control Experiment Significance of the difference Total number of flies investigated 5571 6590 Mean number of progenies per female 99. Table 3.0069 Recombination frequency of the cv and v markers 20.67. genotype D2/D3: F = 3.49 ± 0.05.71 t = 14. the result of values mentioned was calculated according to the Student’s which was as follows: t = 3. the result of which was as follows: t = 3. whereas that of the ex. As shown in control and experimental crosses are given in (Figure 4). genotypes in the upper right corner of the figure. 25˚C and 29˚C.35.0007.0007). P = 0. viz.Testing types is also given and indicated by the red diamond (♦). ance. The mean frequency together with its standard deviation for both geno. df1 P = 0. df2 The significance of the difference of the mean values mentioned =56.84 37. P = 0.67. df = 12159.48. df2 = 53. the frequency in the control dependent on the temperature.61 t = 5. Test- of the heterogeneity of the results inside each mus309 genotype ing of the heterogeneity of the results inside each mus309 was carried out using the one-way analysis of variance. frequency together with its standard deviation for both geno- types is also given and indicated by the red diamond (♦). P < 0.42 ± 0.47 t = 3.

However. The mean frequency ferences are the same as those calculated on the basis of together with its standard deviation for both genotypes is also the mean values per female or per culture bottle. genotype D2/D3: F = 2. df1 = 2. The columns represent the mean fre- given in (Figure 7). in each temperature for the control and experimental crosses are given in (Figure 5).095. df2 = 56. As appears from the figure. df1 = 2. The effect of temperature on the recombination fre- distance in the experimental crosses was significantly quency of the v and f markers of the X chromosome in the prog- higher than in the control crosses (P < 0. 18˚C. ods of the comparison of the mus309 genotypes studied. The significance of the difference of the mean values men. df = 114 and P = lows: None of the parameters measured was dependent 0.0001.0001). the P-values for the test of heterogeneity are given below the group of columns for the respective geno- type. temperature for the control and experimental crosses are 18˚C. OJGen . The results mental than in the control crosses (P < 0. The significances of the dif- vertical lines their standard deviations. the map quencies of recombinant of progenies per culture bottle and the distance in the control crosses was not dependent on the vertical lines their standard deviations. and that for the test of the difference between the geno. whereas that in the experimental crosses together with its standard deviation for both genotypes is also was. the two meth- 0. tioned was calculated according to the Student’s t-test.86. However. 25˚C and 29˚C.0002. It also appears that the mean value of the frequency in question was signify- cantly lower in the experimental than in the control crosses (P = 0. were as follows: genotype +/+: F = 0. Testing of the heterogeneity of the results inside each mus309 genotype was tance in question was significantly higher in the experi- carried out using the one-way analysis of variance. viz. essentially. However. P = 0. the In general. The coefficients of coincidence in each temperature for the control and experimental crosses are given in (Fig- ure 8). except that the comparison of the recombi- heterogeneity of the results inside each mus309 genotype was carried using the one-way analysis of variance.72. i. their map distances. directly on the basis of the total numbers of progeny in 25˚C and 29˚C. their map distances. df2 = 56. and that for the test of the difference between the geno- types in the upper right corner of the figure.39. P = 0. the result of which was as follows: t = 5.19. The recombination frequencies of the v and f markers. The columns represent the mean frequencies of the control and experimental crosses and their compari- recombinant progenies in percentages per culture bottle and the sons are given in (Table 3).075. It also appears that the mean value of the map dis. df1 = 2. the mean value of the map Figure 6. gave similar results. its mean value in the experimental crosses was significantly higher than in Figure 5. eny of the mus309+/mus309+ (+/+) and mus309D2/mus309D3 The map distances of the cv and f markers in each (D2/D3) females raised in three different temperatures.0028. the P-values for the test of heterogeneity are given below the group of columns for the respective geno. quency of the cv and v markers of the X chromosome in the prog- The values of all the parameters measured calculated eny of the mus309+/mus309+ (+/+) and mus309D2/mus309D3 (D2/D3) females raised in three different temperatures. the main features of the results are as fol- result of which was as follows: t = 3. the frequency in the control crosses was not dependent on the temperature. df = 115 and P < 0. Portin / Open Journal of Genetics 1 (2011) 38-47 43 The recombination frequencies of the cv and v mark- ers.e. df2 = 53. The effect of temperature on the recombination fre. P = 0. given in given and indicated by the red diamond (♦). The coefficient was not dependent on the tempera- ture in either group of crosses.0001).97. given and indicated by the red diamond (♦). in each temperature for the con- trol and experimental crosses are given in (Figure 6). i. df1 = 2. whereas that of the experimental crosses was. viz. The significance of the difference of the mean values men- tioned was calculated according to the Student’s t-test. P. The results nation frequencies of the cv and v markers gave no sig- were as follows: genotype +/+: F = 2. In the experimental crosses types in the upper right corner of the figure.0001). the control crosses (P < 0.45. P = nificant difference. genotype D2/D3: F = 6. In addition.e. df2 = 53.56. Copyright © 2011 SciRes. mutant mus309 females. The mean frequency temperature. Testing of the the figures. In addition. As illustrated in the figure. involving mutant mus309 females all the parameters.0002). The frequency was temperature sensitive in neither group of crosses. on the temperature in the control crosses involving non- type.

df = 114 and P < 0. is interesting. failure of the repair of the DSBs. 18˚C. the result of which was as The results were as follows: genotype +/+: F = 1. It is logical to assume that of Drosophila melanogaster affecting crossing over the formation of the DSBs and their subsequent repair is which also affect interference involve preconditions of temperature-dependent in the mus309 mutants. P = 0. according to the Student’s t-test. except the frequency of true single crossovers in the cv-v and that for the test of the difference between the genotypes in interval.0001. It is. These ence based on the genetic distance. known that a defect in DSB repair activates a signaling in all the parameters studied. as the results suggest (Figure 1). df2 = 56. DISCUSSION the crossing over frequencies. Consequently. and that for the test of mentioned was calculated according to the Student’s t-test. 25˚C centimorgans (cM) per culture bottle and the vertical lines their and 29˚C.0001. df1 = 2.97. the = 53.21. viz. df = 12160 and P < of the figure. for both genotype +/+: F = 1. the P-values for the test of heterogeneity are given below the group of columns for the respective genotype. df1 = 2. but are in contradic- ideas are also consistent with the suggestions which will tion with the models based on physical distance. df1 = 2.44 P. 18˚C. and the coefficient of coincidence. The significance Testing of the heterogeneity of the results inside each mus309 of the difference of the mean values mentioned was calculated genotype was carried out using the one-way analysis of variance. The mean value of C calculated on the basis cated by the red diamond (♦). The significance of the difference of the mean values of columns for the respective genotype. The columns represent the mean coefficients of co- standard deviations. the the difference between the genotypes in the upper right corner result of which was as follows: t = 8. 4.20.2.1. were dependent on the temperature. The results were as follows: according to the formula given in material and methods. it is also logical to assume plained in the following discussion.63.58. D2/D3: F = 6. in fact. as compared to the It is convincingly established that those meiotic mutants control females. It is therefore suggested that over without affecting interference involve the crossing the decrease in the fertility observed is due to a complete over event itself [36]. Portin / Open Journal of Genetics 1 (2011) 38-47 Figure 7. these results support that failure of the repair is the more complete the higher the counting number model of the mechanism of interfer- the temperature. viz. df2 = 53. genotype D2/D3: F = 1.0038. 0. standard deviations. whereas those mutants that affect crossing are deficient in such repair. P = P-values for the test of heterogeneity are given below the group 0. genotype genotypes is also given and indicated by the red diamond (♦). except for the frequency of pathway that leads to defects later in the oocyte devel- the true single crossovers in the cv-v interval. which was calculated one-way analysis of variance. thus leading to sterility died. OJGen . which crossing over.65. the upper right corner of the figure. df1 = 2. The mean map distance together with its incidence (C) per culture bottle and the vertical lines their standard deviation for both genotypes is also given and indi. As ex- opment [32-35]. Testing of the heterogeneity of the of the total number of progeny of the females of the genotype results inside each mus309 genotype was carried out using the given together with its standard deviation. The effect of temperature on the map distance be- tween the cv and f markers of the X chromosome in the progeny Figure 8. the genes involved Copyright © 2011 SciRes. The effect of temperature on the crossover interfer- of the mus309+/mus309+ (+/+) and mus309D2/mus309D3 (D2/D3) ence in the cv-v-f region of the X chromosome in the progeny of females raised in three different temperatures. In addition.29.16. The columns represent the mean map distances in females raised in three different temperatures. Moreover. P = 0. df2 = 56. the recombination frequency of the v and f ma. When comparing the genotypes stu. df2 follows: t = 7. The Effect of the Temperature and the Mutation on Fecundity mus309 Gene on Crossing Over and Crossover Interference The decrease in and temperature sensitivity of the fe- cundity of the experimental females. 25˚C the mus309+/mus309+ (+/+) and mus309D2/mus309D3 (D2/D3) and 29˚C. be presented below on the effect of the temperature on 4. P = 0.28. In addition. rkers. a significant difference between them was observed of the female or lethality of its progeny [13]. The Effect of Temperature and the mus309 4.

as if the meiocyte was able to ‘count’ dx 2 d recombination events. It is logical to crossing over. the mus309 mutations affect both into a crossover point. ferent categories. One of the models belonging to this category. accordingly. In general. pair-annihilation of the random walkers. such as temperature in the present study. if a mutant that acts on ence is dependent on physical distance (microns or base crossing over also affects interference. neighboring Cx’s must have a certain number. Let d be the probability of the ful. models of crossover This seems. is conclude that interference decreases if the α value in- the repair of DSBs—a necessary condition for crossing creases and/or h decreases [40]. it must influence pairs) between the adjacent crossovers. is between them. 4. it is hard to conceive. but In contrast. walking” precursor becomes immobilized and matures served in this study. the other models are qualitative. Therefore. called the “counting model” [4. crossover interference based on physical distance on the ent on genetic (i. but sors. ing-over frequencies. As mentioned in the introduction. a “random As has been shown earlier [25. rently only one model. In particular. initial density of the random walkers. The central feature of the counting model is that re- ing over in one region and only in that region in a combinational intermediates (C’s) have two fates – they three-point crossing-over experiment.e. What in this respect is true for meiotic mutants is. m. sent study are consistent with this idea. α. P. To my knowledge. is quantitative while course. h. however. overs. According to the reaction-diffusion model. which the mus309 gene product affects. also true for other factors that affect crossing over. however. In the control crosses. viz. in terms of suggests that temperature affects the formation of DNA the reaction-diffusion model. C  (3) The second category of models. i. decrease in mus309 mutants [25]. however. the initial density of crossover precur- temperature affects the event of crossing over itself. This indicates that factors only. while non-crossovers the m value of the counting model should the DSBR pathway remains functional. hypothesizes that crossover interfere- Since C is independent of x. re. assumes that interference is depend. are not compatible with the reaction-diffusion model. over. This crossovers. The first category of models. i. Portin / Open Journal of Genetics 1 (2011) 38-47 45 are called precondition genes and exchange genes. Let b be the prob. DSBs. and the rate of their processing into only in combination with the mus309 mutation. to be the case in the results of the interference can. As indicated in the introduction. meaning that they act prior to the time when mobilized point. or by annihila- the class that has been referred to as the “precondition tion of a random walker due to its collision with an im- mutants”. i. interfere.e.3. The interference is caused by a crossing over and crossover interference and. probability of the fulfillment of preconditions of cross. This model has two parameters—the crossovers are actually generated [39]. the precondition of of their processing into crossover points. the target of the effect is the along the bivalent and determines the distribution of exchange itself [37]. OJGen . called the A involve some precondition of crossing over belonging to particles. of the reaction-diffusion model [40]. the map distances. between adjacent crossovers. Test of the Models of Crossover Interference would not. The results of the pre- In the experimental crosses temperature affected cross.e. in principle. it is also clear that in the day junctions will be resolved as crossovers instead of mus309 mutants the SDSA pathway is blocked. linkage map) distance (Morgans) following grounds: the map distances in the experimen- Copyright © 2011 SciRes. cur- spectively. due to their collision together. be divided into two dif. C.5]. would change due to the effect of temperature but their distances. called The results are also in contradiction with any model of genetic models [4].38]. models suggest that some kind of physical signal travels ence remains unaltered. given the pre. present study. as the initial density of DSBs does not change. According to this model. how the number of cross- double-strand breaks. and interference results from constraints on the fillment of the preconditions in both regions at the same resolution of C’s. however. the results of the present study it did not affect interference. and was also ob. can be resolved with crossing over (Cx) or without (Co). it is known that the MUS309 protein It is also quite logical to assume that if the mus309 is involved in the SDSA pathway of the repair of the mutations affect the balance by which the double Holli- DSBs. ability of fulfillment of the same in another region and The C’s are distributed at random with respect to each only in that region. Then the coefficient of coincidence. crossovers. the double crossovers included. and x the probability of exchange. which may be called x  a  d  x  b  d   a  d  b  d  physical models. neither crossing over frequencies nor interfere. interference depends on two ence was affected by the temperature. of Co’s conditions. and the rate. If. More specifically. other. This was theoretically shown as follows: Let a be the falls into this category. The basic constraint is that each pair of time.e. these the preconditions of crossing over. interference.

On the other hand...072902. and Symington. defective in DNA sis-specific DNA double-strand breaks are catalyzed by double-strand break repair.P. doi:10.C. and Hickson. van Stan. (2006) Initiation of meiotic series of crosses. doi:10. Harris. 14. 50. 409-459.M. [2] Muller.043 the language.J. [22] Brabant. [4] Foss. J.. As explained above. M.13. and Kleckner. and Steinberg. S. a member of a widely conserved protein family. and Lindsley.D. (2009) Drosophila PCH2 is required for a pachytene checkpoint that monitors dou- if interference was dependent on physical distance. (1975) Differential sensitivities and the sophila BLM in double-strand break repair by synthe- target of heat-induced recombination at the base of the X sis-dependent strand annealing. 28. N. However.genet.K. doi:10.2003.. (1915) A linkage variation in Drosophila. ACKNOWLEDGEMENTS 233-271. 375-384. A. 548-557. K. R. Lennon.. (2004) Crossover interference. PhD helped me by conducting sight? Trends in Biochemical Sciences. 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