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Russell
CHAPTER 15
Non-Mendelian Inheritance
Chapter 15 slide 1
2. Many proteins of both mitochondria and chloroplasts are encoded by nuclear genes, indicating that genes have moved from the organelles to the nuclear DNA.
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b. GC content of mtDNA often differs from nuclear DNA, allowing separation by CsCl density gradient centrifugation.
c. Mitochondrial DNA lacks histone-like proteins. d. Multiple genomic copies are in multiple nucleoid regions within the mitochondrion. e. Gene content is conserved across species, but size of the mtDNA varies widely:
i. Animal mtDNA is less than 20 kb (human is 16,569 bp). Essentially all of this DNA encodes products.
ii. Yeast mtDNA is about 80 kb, and not all of this DNA encodes products. iii. Plant mtDNA ranges from 100 kb to 2 million base pairs. Not all encodes 601 20000 Chapter 15 slide 3 products.
Fig. 15.2 Model for mitochondrial DNA replication that involves the formation of a D loop structure
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
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Chapter 15 slide 4
3. Replication of mtDNA is semi-conservative, uses mitochondrial DNA polymerases and RNA primers, and involves no proofreading.
a. Replication of mtDNA occurs throughout the cell cycle. (Contrast with nuclear DNA, which replicates only in S phase.) b. Both strands of mtDNA in most animals replicate in a continuous manner, with replication of one strand initiating well before the other.
4. Mitochondria are not synthesized de novo, but arise from growth and division of preexisting mitochondria (Luck, 1963). Some of their genetic information is in the mitochondrial chromosome, the rest in the nuclear DNA.
a. The mtDNA map (Figure 15.3) contains information for: i. tRNAs. ii. rRNAs. iii. Some polypeptide subunits of cytochrome oxidase, NADHdehydrogenase, and ATPase. b. Nuclear DNA encodes other mitochondrial components, including: i. DNA polymerase and other replication proteins. ii. RNA polymerase and other transcription proteins.
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
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Chapter 15 slide 6
5. Mitochondrial ribosornes translate mRNAs from the mitochondrial chromosome within the organelle. For example, human mitochondrial ribosomes:
a. Have two subunits, 45S and 35S, forming a 60S mitochondrial ribosome. b. Have only two rRNAs, 16S rRNA in the large subunit, and 12S rRNA in the small subunit. c. Usually have only one gene for each rRNA in the mtDNA.
d. Ribosomal proteins are usually encoded by nuclear DNA, and move into the mitochondria from the cytoplasm.
6. Mammalian mtDNA is transcribed into a single large RNA molecule that is cleaved to produce mRNAs, tRNAs and rRNAs. These RNAs are then processed:
a. mRNAs receive 3 poly(A) tails. b. tRNAs receive 3 CCA sequences. c. Mitochondrial mRNAs have no 5 caps.
7. The much larger mitochondrial genomes of yeast and plants differ from animal mitochondria:
a. tRNA genes do not separate genes, and other DNA sequences signal transcription termination. b. Large gaps occur between genes in the mtDNA. c. Introns are found in mitochondrial genes from these organisms, but never in animal mtDNA genes. d. mtDNA sequences encoding some mRNAs do not include a complete stop codon. Instead, the 3 end is U or UA, and the poly(A) tail completes the stop codon (UAA).
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i. Yeast and plant mRNAs have a 5' leader sequence, and so initiation can occur at the first AUG codon.
ii. Animal mitochondria lack the leader sequence, and so initiation must occur in a unique way. b. In translation, mitochondria have similarities to bacteria:
d. Mitochondria show extended wobble in base pairing of tRNA with codon, allowing translation with only 22 tRNAs (compared with 32 tRNAs theoretically needed for translation using standard wobble).
e.. Structural features that never differ in cytoplasmic tRNAs sometimes do differ in mitochondrial tRNAs. 601 20000 Chapter 15 slide 8
Fig. 15.4 Synthesis of the multisubunit protein cytochrome oxidase takes place on both cytoplasmic and mitochondrial ribosomes
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
601 20000
Chapter 15 slide 9
ii. The woman's mtDNA did not match the mtDNA of living members related through maternal lines. Thus, she was not Anastasia.
d. Conservation biology also uses mtDNA analysis to determine genetic variability in populations (e.g., grizzly bears in Yellowstone Park).
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Chloroplast Genome
1. Chloroplasts have a double membrane, internal lamellar structure containing chlorophyll, and protein-rich stroma. Chloroplasts divide and grow in the same way as mitochondria. 2. The chloroplast genome (cpDNA) is not as well characterized as mtDNA, but some things are known:
a. Structurally, cpDNA is similar to mtDNA. It is dsDNA, a super- coiled circle lacking structural proteins. b. The GC content of cpDNA often differs from both nuclear and mtDNA, allowing separation on CsCl density gradients. c. The size of cpDNA varies from 80 kb-600 kb. All chloroplast genomes carry noncoding DNA.
d. Each chloroplast has multiple copies of cpDNA in several nucleoid regions. An example is Chiamydomonas with 500-1,500 cpDNA molecules per chloroplast.
3. Nuclear genes encode some chloroplast components, while cpDNA genes (which may include introns) encode the rest, including (Figure 15.5):
a. Two copies of each chloroplast rRNA (loS, 23S, 4.5S and 5S). The copies are included with other genes in inverted repeats designated IRA and IRB that define the short (SSC) and long (LSC) single copy regions of the cpDNA. b. The tRNAs (30 in tobacco and rice, 32 in the liverwort Marchantia). c. Almost 100 highly conserved ORFs. Approximately 60 are now correlated with proteins needed for chioroplast transcription, translation aiid photosynthesis. 601 20000 Chapter 15 slide 11
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
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Chapter 15 slide 12
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Chapter 15 slide 14
2. Results of crosses between plants with shoots that are variegated illustrate this phenomenon (Table 15.2):
a. When ova are from green plants, only green progeny result, regardless of pollen source. b. When ova are from white plants, only white progeny result (but soon die from lack of chlorophyll), regardless of pollen source. c. When ova are from variegated plants, all three types of progeny result, regardless of pollen source.
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3. Shoot color in these plants therefore shows a pattern of maternal inheritance. There are three assumptions in the model:
a. Pollen contributes no chloroplasts or leucoplasts to the zygote. b. The chloroplast genome replicates autonomously, so that progeny plastids retain the same color phenotype as the original plastid. c. Segregation of plastids during eukaryotic cell division is random, providing some offspring cells with chloroplasts, some with leucoplasts, and some with a mixture.
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Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
601 20000
Chapter 15 slide 17
Fig. 15.7 Model for the inheritance of shoot color in the four oclock
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
601 20000
Chapter 15 slide 18
2.
Experimental crosses in Neurospora involve fusion of nuclei from mating type A and a parents. Crosses can occur two ways:
a. b. Place both parents on medium at the same time. Inoculate one parent onto medium, and add the second parent several days later. The first parent produces all the protoperithecia (fruiting bodies containing the ascospores).
3.
e.
The [poky] allele results from a 4-bp deletion in the promoter for 19S rRNA in the mtDNA, resulting in deficiency of small ribosomal subunits and greatly decreased mitochondrial protein synthesis.
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Fig. 15.8 Results of reciprocal crosses of [poky] and normal (wild-type) Neurospora
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
601 20000
Chapter 15 slide 20
b. The spontaneous mutation rate is 0.11%, but exposure to an intercalating agent (e.g., ethidium bromide) raises the rate to 100%.
c. This allows isolation of different petite cell lines, containing different mutations.
3. Yeast crosses between petite and wild-type cells (a X crosses) determine the mechanism of inheritance for this phenotype.
a. The zygote formed from mating is grown into a colony to check its phenotype, and when it sporulates by meiosis, the tetrad of ascospores can also be grown into colonies for phenotype analysis.
b. Some petite X wild-type crosses give 2:2 segregation (wild-type:petite). i. This is the same ratio as seen in nuclear genes, so these petite mutants are nuclear (segregational) petites, written pet- (Figure 15.9). ii. The cross in this case was pet- X pet+. Diploid was pet-/pet+ (hence wild-type) and the spore tetrad contained 2 pet- and 2 pet+ spores.
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Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
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Chapter 15 slide 22
c. Another class of petite mutants is the neutral petites ([rho-N]). i. When crossed with wild-type ([rho-N] X [rho+N]) produce wild-type diploids ([rhoN]/[rho+N]) and spores that segregate 0:4 (no petite : 4 wild-type). ii. This is an example of uniparental (not maternal, since gametes are same size) inheritance. iii. In [rho-N] mutants, nearly 100% of the mtDNA is missing, and so mitochondrial functions are also missing. iv. Spores produce only wild-type colonies because normal mitochondria from the wildtype parent provide normal mitochondria for the progeny. The petite trait thus is lost after one generation. d. Most petite mutants are of the suppressive ([rho-S]) type. They differ from neutral petites by having an effect on the wild-type, although both are mutations in mtDNA. i. A [rho+/rho-S] diploid has a respiratory-deficient phenotype, and if it divides mitotically the progeny will nearly all be petites. ii. Sporulation of the petite [rho+/rho-S] diploid produces tetrads with a 4:0 (petite : wildtype) ratio. iii. Sporulation of the rare wild-type [rho+/rho-S] diploid produces tetrads with a 0:4 (petite : wild-type) ratio. iv. Suppressive petite mutants start with deletions in mtDNA. The amount of mtDNA is restored by duplications of existing mtDNA, often creating gene deletions and rearrangements that cause deficiencies in the enzymes for aerobic respiration. v. The suppressive effect over normal mitochondria might result from either: (1)Faster replication of the mutant mitochondria, outcompeting wild-type, or (2)Fusion with normal mitochondria and recombination between [rho -S] mtDNA and 601 20000 Chapter 15 slide 23 wild-type mtDNA.
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
601 20000
Chapter 15 slide 24
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
601 20000
Chapter 15 slide 25
2.
A chloroplast-encoded trait in Chlamydomonas is erythromycin resistance ([eryr]), with wild-type cells sensitive to the antibiotic ([erys]).
a. c. The cross mt+[eryr] 3 mt-[erys] produces about 95% erythromycin resistant progeny (thus uniparental inheritance). In both crosses, progeny resemble the mt+ parent, reflecting either i. Biased segregation of one chloroplast type, or ii. Inactivation of one chloroplast type. b. The cross mt-[eryr] 3 mt+[erys] produces about 95% erythromycin sensitive progeny (also uniparental inheritance).
3.
Other Chlamydomonas chloroplast alleles also show uniparental inheritance, with the progeny phenotype always the same as the mt+ parents phenotype.
a. Density labeling experiments with mt+ and mt- cpDNA show that cpDNA from the mt- parent disappears in mt+/mtzygotes.
4.
While 95% of progeny show uniparental inheritance, the remaining 5% display biparental inheritance, with both types of cpDNA present and active.
a. c. The genetic condition of these zygotes is called cytohet (cytoplasmically heterozygous). Occasionally, a biparental zygote retains both traits in subsequent generations. This results from recombination in the cpDNA. b. Cytohet cells usually produce pure types on successive mitotic divisions. (This is similar to variegation in Mirabilis.)
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Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
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Chapter 15 slide 27
ii. Abnormal pigment accumulation on the retina, causing chronic inflammation and degeneration of the retina.
iii. Heart disease. iv. Kearns-Sayre syndrome results from deletions in mtDNA. A model for the disorder is that tRNA genes are removed, disrupting mitochondrial translation.
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c. Myoclonic epilepsy and ragged-red fiber disease (MERRF). Symptoms include: i. Microscopic tissue abnormality, ragged-red fibers. ii. Myoclonic seizures (jerking spasms). iii. Ataxia (uncoordinated movement). iv. Accumulation of lactic acid in blood. v. Additional symptoms are sometimes present, including: (1)Dementia.
(2)Loss of hearing.
(3)Difficulty speaking. (4)Optic atrophy. (5)Involuntary jerking of eyes. (6)Short stature.
2. In most mtDNA disorders, cells of affected individuals have a mix of normal and mutant mitochondria (heteroplasmy).
a. Proportions of the two mitochondrial types vary between tissues, and between individuals.
b. Severity of disease correlates with the relative amount of mutant mitochondria.
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d. Transmission of these viruses between yeast cells occurs in mating. All progeny of the mating inherit copies of the parental viruses.
2. Killer yeast cells are an example of an infectious mechanism of cytoplasmic inheritance. 601 20000 Chapter 15 slide 31
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(1) The F2 genotypes also have a 1:2:1 ratio (D/D : D/d : d/d). All F2 snails have sinistral shells.
(2) Selfing the F2 produces an F3 that is all dextral, due to the D/d genotype of the F2 mothers (who had sinistral shells).
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Fig. 15.14 Maternal effect: Inheritance of the direction of shell coiling in the snail Limnaea peregra
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
601 20000
Chapter 15 slide 35
4. In the snail shell example, direction of coiling is determined by the orientation of the mitotic spindle in the first mitotic division following fertilization. Maternal products within the oocyte direct orientation of the mitotic spindle, and thus shell coiling.
a. This is supported experimentally: i. When eggs of d/d mothers are injected with cytoplasm from dextral snails, dextral progeny result. ii. When eggs of D/_ mothers are injected with cytoplasm from sinistral mothers, the progeny still have dextral shells. b. Interpretation is that: i. The D allele produces a cytoplasmic product that causes dextral coiling.
ii. The d allele does not produce this product, and sinistral coiling is produced by default.
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Genomic Imprinting
1. Most nuclear genes function independently of maternal or paternal origin. The expression of some nuclear genes, however, is determined by genomic imprinting (parental imprinting), where one parents allele is expressed preferentially. 2. Human examples of genomic imprinting include:
a. Prader-Willi syndrome (PWS): i. Affected individuals are small and weak at birth, with retardation and poor feeding. ii. Feeding difficulty resolves into uncontrollable eating and associated problems that are typically fatal by age 30. iii. The cause is a disruption in chromosomal region 15q11-q13. (1) In 7080% of cases, the disruption is on the fathers chromosome 15. (2) In PWS patients, the maternal 15q11-q13 region is normally suppressed by methylation of the genes. Paternal genes are needed for normal development, and when they are disrupted PWS results. b. Angelman syndrome (AS) produces severe motor and intellectual retardation, small head, jerky movements, hyperactivity, and unprovoked laughter. i. About 50% of AS patients have a deletion of region 15q11-q13. ii. In AS, maternal alleles are needed for normal development, because paternal genes are inactivated by methylation prescribed by genomic imprinting. Defective maternal genes result in disease.
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