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L 8: Epistasis, Complementation
and Biochemical Pathways
Epistasis is used in 3 different ways, variations
on interactions between genes:
(1) classical epistasis: how an allele of one gene
blocks or masks the effect of an allele of another
gene, altering classical expected ratios
(2) functional epistasis: relationships between the
expression of different genes (enzymes) operating
in the same pathway
(3) statistical epistasis: a nonadditive effect in
quantitative genetics
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(3) Dominance explained by biochemical interactions: Nonfunctional mutations are recessive, haplosufficient alleles are
dominant. Note there are 2 alleles affecting dominance in the
heterozygote:
(1) functional or haplosufficient allele, (2) non- functional allele
Scale distinctions
dominance, incomplete dominance and codominance are somewhat
arbitrary. these categories are designed for the convenience of
analysis.pp 215
This could suggest to you that the authors think classical analysis models are
arbitrary, but what they are saying is genetic analysis, must :
(1) define the scale of analysis and (2) the scale of the causal mechanisms.
Thus, (1) dominance, incomplete dominance and codominance are
observations, empirical explanations that can be repeatedly confirmed
by careful breeding studies. These categories are used in classical
models to interpret visible phenotype ratios, signifying underlying
genotype ratios and a class of allelic interactions.
But a mechanistic explanation of dominance on a phenotypic level may
involve:
1) biochemical interactions (physiology) 2) haplosufficiency (level of gene
expression), or 3) modifier genes (gene interaction).
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10
X
or
A
F1
Same (ww)
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different
(complementation)
X
or
F1
Same (ww)
11
Gene W1+ ,
Gene W1+ ,
Gene w2
-
Gene w2-
They complement
Gene W1+ . Gene w2-
Gene w1- . Gene W2+
(W1+/W1+, w2-/w2-)
(w1-/w1-, W2+/W2+)
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9 W1+__ W2+__
3 w1-w1- W2+__
3
1
W1+__ w2-w2-
w1-w1- w2-w2-
genes.
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recessive epistasis
without a cumulative
effect
7
(2) Independent chromosome
assortment but an epistatic
interaction
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Gene w1-
Gene W2+
Gene w1-
Gene W2+
Enzyme 1
Compound A
(white)
Enzyme 2
Compound B
(white)
Compound C
Gene W1+
Gene W1+
Gene w2-
Gene w2-
Enzyme W1+
Compound A
(white)
Enzyme2
Compound B
(white)
Compound C
(blue- not expressed)
White 2
Gene W1-
Gene W2-
Gene W1+
Gene W2+
Enzyme 1
Compound A
(white)
Compound B
(white)
Enzyme2
Compound C
(blue- expressed)
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# of genes ?
19
20
21
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(1)Induce mutation
variation, make
heterozygotes, promote
haplotype spore
variation..
(2) Screen on minimal media for
nutritional mutants (no growth).
(3) Select for arginine requiring
mutants of Neurospora from the
original haploid individuals
inoculated on complete media
Arginine - deficient mutants
grow when arginine (one of
many amino acids) is added to
minimal media, or arginine
rescues these individuals.
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arg-
Some
mutants
could grow when
related compounds
were added to the
media
instead of arginine,
suggesting a
biosynthetic
pathway.
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Supplement
arg-1+
Enzyme 1
precursor
arg-2+
Enzyme 2
ornithine
arg-3+
Enzyme 3
citrulline
arginine
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precursor
ornithine
citrulline
arginine
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Genetic Analysis of a Metabolic Pathway - Through a trial by sampling and best chemical
guess process, four compounds appear required to make compound 4, this is a measurable
product, a metabolic character or phenotype. You have also found by testing different
bacterial populations there are mutants (eg B) for specific enzymes catalyzing a specific
compound transformations. Rather than beginning a long biochemical analysis, you can use a
genetic analysis of the mutations and participating compounds to order the metabolic pathway
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(+) indicates that the product phenotype (wild type) was expressed. (-) indicates the phenotype
has not been produced or expressed, indicating an enzyme mutant.
(1)What is the order of supplement compounds (1-6)-look at the columns
Start here- the supplement at the beginning of the pathway never rescues the pathway. The final
supplement in the pathway always rescues the pathway.
(2) What is the order of enzymes (A-E) in the pathway -look at the rows
Start here- which enzyme catalyzes the first reaction means which mutant enzyme is rescued by
all but 1 compound - which must be the precursor substrate. One mutant enzyme is only rescued
by 1 compound , indicating it is at the end of the pathway.
Supplement
1
2
3
4
5
-
-
-
-
+
+
+
+
+
+
Enzyme
A
B
C
D
E
-
+
-
+
+
-
-
-
+
+
+
+
-
+
+
-
-
-
-
-
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1
2
3
4
5
6
Enzyme
A
B
+
+
+
+
-
+
-
+
-
+
-
+
C
D
E
-
+
+
+
-
+
-
-
-
-
-
+
-
-
-
+
-
+
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