Worm Breeder's Gazette 11(2): 105

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

Dominant Negative Mutations of let-60 Define its Controlling Role in Vulval Induction

Min Han, Raffi Aroian and Paul Sternberg

Figure 1

We reported previously the characterization of 7 dominant vulvaless (
Vul) mutations, isolated as dominant suppressors of a lin-16 
multivulva (Muv) mutation [WBG 10 (3): 132].  These mutations, six of 
which are recessive lethal, were found to be alleles of let-60, 
previously identified by recessive lethal alleles [Clark et al., 
Genetics, 119: 345 (1988)].  We refer to these semi-dominant alleles 
as let-60(sd).  The conclusions from our further studies on let-60 can 
be summarized as follows.
let-60 and lin-34 are likely to be the same gene.  let-60(sd) 
alleles were mapped between dpy-20 and the left break-point of nDf27.  
lin-34, identified only by semi-dominant Muv mutations, maps between 
dpy-20 and let-60 (G.  Beitel, G.  Jongeward, per.  comms; our data).  
Our mapping data suggest that both are about 0.01 mu to the left of 
dpy-20.  Also, lin-34 suppresses in trans the let-60(sd) Vul phenotype.
In fact, we have isolated a lin-34(sd) allele as a dominant 
suppressor of the dominant suppressor phenotype of let-60(sd).  More 
intriguingly, the lethality of some let-60(sd) homozygotes can be 
maternally rescued by lin-34/+.Vulvaless is a loss-of-function 
phenotype of let-60.  We have isolated two intragenic revertants of 
let-60(sd) by reverting the dominant Vul or suppressor phenotype of 
let-60(sd).  These revertants and the three previously isolated 
alleles (Baillie's lab) are likely null mutations because they behave 
similar to Dfs in heterozygotes.  let-60(null)/let-60(sd) animals from 
a lin-34/let-60(sd) mother show a more severe defect in vulval 
induction (0%) than let-60(sd)/+ (57% for the allele used), indicating 
that Vul is a loss-of-function phenotype.  Thus, because Df/+ has 
wildtype vulva, let-60(sd) alleles are dominant negative mutations.
The level of let-60 activity controls VPCs fates.  In let-60(sd)/+ 
animals, let-60 activity is lower than that in let-60(null)/+, 
resulting in a Vul phenotype.  We propose, in light of our argument 
that lin-34(sd) are alleles of let-60, that a lin-34(sd) mutation 
results in elevation of let-60 activity and consequently a gonad 
signal independent Muv phenotype.  Thus, a high level of let-60 
activity specifies 1  or 2  while a low level specifies 3 .  It is 
also conceivable that lin-34(sd) may be mutations in a distinct gene 
located very close to let-60, but nonetheless result in an increase of 
let-60 activity.
Genetic interactions of let-60/lin-34 with other genes suggest its 
position in the vulval induction pathway.  The following pathway was 
deduced from examination of double mutant 
phenotypes:
[See Figure 1]

Figure 1