Worm Breeder's Gazette 11(2): 118

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dpy-29 and tra V Mutations Appear to Define a Single Locus that Coordinately Regulates Sex Determination and Dosage Compensation

Leslie DeLong, John Plenefisch and Barbara Meyer

Figure 1

The cumulative genetic evidence continues to support the proposal 
that dpy-29, like sdc-1 and sdc-2, is required in XX animals for the 
proper control of both sex determination and dosage compensation.  
However, unlike the sdc genes, the dosage compensation and sex-
determination functions of dpy-29 appear to be independently mutable.  
Furthermore, null or near null alleles of dpy-29 do not normally 
masculinize XX animals, rather, the cryptic Tra phenotype can only be 
seen in certain genetic backgrounds.  The tra V mutations, which cause 
recessive masculinization of XX animals, appear to result from loss of 
only the sex-determination function of the gene.  One dpy-29 allele 
appears to disrupt only the dosage-compensation function, the 
remaining dpy-29 alleles appear to disrupt both gene functions.
dpy-29 is required for the XX mode of dosage compensation.  
Mutations in dpy-29 result in recessive XX-specific dumpiness and XX-
specific maternal effect lethality; the lethality ranges from 72% to 
98%.  Phenotypically, mutations in dpy-29 resemble mutations in the 
dosage compensation dpy genes dpy-26, 
he lin-14 assay we have shown 
that mutations in dpy-29 result in elevated levels of X-linked gene 
expression.  Like other mutations that disrupt the XX mode of dosage 
compensation, mutations in dpy-29 rescue xol-1 XO animals.
dpy-29 is required for hermaphrodite sexual development.  dpy-29 
differs from the dosage compensation dpy genes in two ways: 1) dpy-29 
mutations, like sdc-1 and sdc-2 mutations, suppress the lethality of 
xol-1 XO animals, resulting in rescued animals that are male.  By 
contrast, dpy-26, s suppress the 
lethality of xol-1 XO animals but result in rescued animals that are 
hermaphrodite.  2) dpy-29 mutations also differs from dpy-26, 
s in their effects on the sexual 
phenotype of 2X:3A animals.  dpy-29(y100) 2X:3A animals, like dpy(+) 
2X:3A animals, are usually male; by contrast, dpy-26, 
imals are feminized.  The 
masculinizing effect of dpy-29 mutations can be seen directly when 
placed in trans to tra(y52).  dpy-29/tra V XX animals range from 0 to 
99% masculinized, depending on the dpy-29 allele.  This 
masculinization is specific to mutations in these two genes; dpy-29 
mutations show no dominant masculinization with mutations in tra-1 or 
tra-2.  tra V mutations show no dominant masculinization with 
mutations in dpy-26, dpy-28, 
nodpy-29 rs to act 
upstream of her-1.  her-1(+) is required for the masculinization of 
tra V XX animals and for the masculinization of dpy-29/tra V XX 
animals.  This result, combined with the observation that dpy-29 
mutations suppress the XO- specific lethality of xol-1 mutations, 
indicates that dpy-29 acts at the same point of the sex-determination 
pathway as sdc-1 and sdc-2.The dosage-compensation and the sex-
determination functions of dpy-29 are independently mutable.  The 
extent of masculinization of the dpy-29 alleles is unrelated to the 
strength of the dosage compensation defect, which is measured by 
extent of the XX-specific maternal-effect lethality.  For example, 
both dpy-29(y144) and dpy-29(y100) have the same level of maternal-
effect lethality (97%), however 99% of dpy-29(y144)/tra V animals are 
masculinized, whereas only 3% of dpy-29(y100)/tra V animals are 
masculinized.
By a cis-trans test, dpy-29 and tra V mutations are in the same gene.
Three independent tra V dpy-29 mutants were obtained by inducing dpy-
29 mutations on a tra V chromosome (two different tra V alleles were 
used).  All three double mutants are phenotypically indistinguishable 
from strong dpy-29 mutations: XX animals are fully hermaphrodite and 
exhibit a strong maternal-effect lethality (96% - 98%).  We have 
evidence that the original tra V mutation is still present in two of 
the double mutants.  For one double mutant, a Tra non-Dpy recombinant 
was obtained after screening 17,000 progeny of tra V dpy-29/+ mothers. 
In another double mutant, the dpy-29 mutation was shown to be 
suppressible by sup-7(st5).  The tra V dpy-29; 
ed animals are pseudomales, in contrast to 
the tra V dpy-29 mutant animals which, when viable, are hermaphrodites.

[See Figure 1]
Mutations in dpy-29 probably represent the null phenotype.  
Mutations in dpy-29 that have strong cryptic masculinizing effects 
resemble a deficiency in trans to tra(y52).  99% of Df/tra progeny of 
Df/tra mothers and 99% of dpy-29/tra progeny of dpy-29/tra mothers are 
fully masculinized.  However only approximately 50% of Df/tra or dpy-
29/tra progeny of tra/+ mothers are masculinized.
The XX masculinization of mutations in tra V is suppressible by XXX 
and mutations in dpy-26, 
erpretation that dpy-29 mutations 
represent the null phenotype of tra V requires that, in dpy-29 mutants,
the dosage compensation defect suppresses the masculinization caused 
by the sex determination defect.  Not only have we shown this to be 
true for dpy-29 as described in the cis-trans test above, but we have 
found that mutations in dpy-26, c 
the effect of dpy-29 mutations by completely suppressing the 
masculinization of mutations in tra V.  That is, homozygous dpy; tra(
y52) XX animals are hermaphrodites.  In a similar vein, we have found 
that him-5 XX animals are dumpy 
hermaphrodites.
The dosage compensation dpy genes also cause a dominant suppression 
of the masculinization of tra(y52).  dpy/+; tra(y52) progeny of dpy/+; 
tra(y52)/+ mothers are frequently hermaphrodite.  For example: 100% of 
dpy-27/+; tra(y52) or yDf10/+; tra(y52) XX animals are hermaphrodite; 
70% of dpy-28/+; tra(y52) + animals are hermaphrodite; and 30% of dpy-
26/+; tra(y52) or 58% of sDf21/+; tra(y52) XX animals are 
hermaphrodite.  Suppression is not seen in the next generation however 
dpy/+; tra(y52) progeny of dpy/+; tra(y52) mothers are fully 
masculinized.  These results are similar to the results described 
earlier for both a deficiency of dpy-29 and dpy-29 mutations in trans 
to tra(y52).
Unexpectedly, we have found that dpy-26, 
se a maternal-effect suppression 
of the masculinization of tra(y52).  dpy/+; tra(y52) progeny of dpy; 
tra(y52) mothers are mostly hermaphrodite.  This is very different 
from the results seen with dpy-29, which shows little or no maternal-
effect suppression of tra(y52).
There is considerable evidence showing that disruption of dosage 
compensation in XX animals is associated with suppression of 
masculinization in sexually transformed animals.  For example, 
mutations in dpy-26,  the 
masculinization of her-1(n695sd) and her-1(y101sd).  In addition, sdc-
1(y67) XX animals become more strongly transformed when the dosage-
compensation defect is suppressed by y63.  Thus, although feedback 
between sex-determination and dosage compensation is not novel, the 
dpy-29 mutations provide a dramatic demonstration of this feedback 
operating within a single locus.

Figure 1