Yeast Extra Practice Problems

1. You are an S. cerevisiae researcher studying the cal1-1 mutation, which causes the
recessive phenotype of increased sensitivity to calcofluor white (Cal

!. Cells carrying this

mutation fail to grow in low amounts of calcofluor white.
You wish to identify extragenic supressors of cal1-1 that restore calcufluor white
resistance to wild"type levels. You mutageni#e your cal1-1 strain, plate onto media
containing a low amount of calcofluor white and loo$ for colonies that are now able to
grow normally (Cal
%
phenotype!. &rom this screen, you isolate seven different putative
suppressor strains, which you name sup1-sup7.
1a. &or each strain, you would li$e to be certain that the suppression phenotype is caused
by a single mutation. 'sing your choice of yeast strains, describe a genetic experiment
that can be performed to test this. Please state the predicted results (genotypes and
phenotypes! if(
i. the Cal
%
phenotype is caused by a single mutation
ii. the Cal
%
phenotype is caused by two mutations, both of which
are re)uired for calcofluor white resistance.
For each strain, cross to the cal1-1 mutant and observe the segregation pattern.
For a single mutation: all resulting tetrads should segregate 2 Cal
R
: 2 Cal
S
For sup1 this would correspond to supX cal1-1 : S!X cal1-1
For two mutations: "here should be tetrads showing phenot#pes corresponding to
!$, %!$ and "", with the ratio depending on the lin&age o' the mutations.
"he mutant supX-1 supX-2 cal1-1 should be Cal
R
but the other ( genot#pes will
result in a Cal
S
phenot#pe.
1b. You want to eliminate the possibility that any of your sup strains are simply revertants
in the cal1 gene. *escribe a cross that will allow you to show whether or not the sup1-
sup7 mutations are extragenic. Please include the predicted results for either case.
Cross each o' the sup mutants to wild-t#pe and anal#)e the tetrads to determine
lin&age. *' the sup mutation is a true reversion #ou should onl# ever see !$ tetrads.
*' it is an intragenic mutation then most will be !$ but occasionall# there will be
%!$ or "". *' the mutation is e+tragenic #ou should see !$, %!$ and "" in ratios
that depend on the lin&age between the cal1 mutation and the sup mutation.
1c. You determine that the sup1-sup7 strains each contain single mutations in genes other
than cal1. +o estimate the number of genes you have identified, you next perform
complementation tests with you suppressors in cal1-1 bac$ground. (,ote( the data in the
table show the phenotype of the diploid generated by mating the two strains.!
cal1-1 cal1-1
sup1
cal1-1
sup2
cal1-1
sup3
cal1-1
sup4
cal1-1
sup5
cal1-1
sup6
cal1-1
sup7
cal1-1 %
cal1-1
sup1
% % % %
cal1-1
sup2
% % %
cal1-1
sup3
% % %
cal1-1
sup4
% % % %
cal1-1
sup5
%
cal1-1
sup6
%
cal1-1
sup7
%
-. indicates Cal

-%. indicates Cal

%
'sing the data in the table, place the sup mutations into complementation groups. /re
there any mutations that cannot be place into a group0 1f so, explain.
sup, is dominant and so cannot be placed in a complementation group.
sup1, sup( and sup- are in one complementation group, sup( and sup. are another
and sup/ is in a group b# itsel'.
1d. +o further characteri#e two of your sup strains, you perform the cross below,
sporulate the diploids, and analy#e 122 tetrads.
cal1-1 sup1 x cal1-1 sup5

Cal
%
Cal
%
Cal
%
Cal
%
Cal

Cal
%
Cal

Cal
%
Cal
%
Cal

Cal
%
Cal
%
3 tetrads( 14 44 15
%!$ "" !$
6hat tetrad classes are the P*, ,P*, and ++0
Provide a brief genetic explanation for the behavior of the sup1 and sup5 strains. Your
explanation should ta$e into account the above lin$age data and the complementation
data presented in part c.
"he above cross indicates that sup1 and sup- are unlin&ed. "his result is une+pected
since the two mutations 'ailed to complement each other. "his is thus a case o'
unlin&ed non-complementation.
7. omeone gives you an S. cerevisiae strain with the unusual property of ma$ing blue
colonies. +o understand this phenotype in greater detail, you cross the blue strain by a
wildtype strain, sporulate the diploids, and dissect tetrads. 1n 82 tetrads, all progeny are
blue9 that is all tetrads segregate :(2 for the blue(normal phenotype.
7a. ;ow do you explain this segregation pattern0
"his segregation pattern is consistent with c#toplasmic inheritance 'or the 'actor
responsible 'or the blue phenot#pe.
7b. You want to identify genes that can modify the blue phenotype. +o do this, you cross
the blue strain by the S. cerevisiae nonessential deletion set and examine the color of the
meiotic progeny. /fter sorting through several candidates and performing additional
tetrad analysis, you have identified two classes of mutants(
class 1 < this deletion set mutant is designated as yfg1::G418R
cross( blue x yfg1::G418R
result( 82 tetrads are analy#ed and all segregate 7 blue =:15 ( 7 normal =:15%
class 7 < this deletion set mutant is designated as yfg2::G418R
cross( blue x yfg2::G418R
result( 82 tetrads are analy#ed and all segregate 2(:or blue(normal and 7(7 =:15(=:15%
Provide a brief explanation for the phenotypes observed for each class of deletion.
"he class 1 deletion is a deletion in a genomic gene that results in suppression o' the
blue phenot#pe. 2 o' the , spores will inherit the deletion. "hese will be white and
0,11R, while the other 2 will remain blue and be 0,11S.
Class 2 deletion arises 'rom a deletion o' a genomic gene, but gives a 2:, segregation
pattern consistent with c#toplasmic inheritance. "his deletion seems to cause a
c#toplasmic product to be made that can suppress the blue phenot#pe. Since 3F02
is deleted, it can4t be a product o' that gene. 5ut, 3F02 could suppress another
gene, so when it is deleted that gene is able to create a c#toplasmic product to
suppress the blue phenot#pe. "he 2:2 0,11S : 0,11R ratio indicates that genomic
$%6 is segregating normall#.