Worm Breeder's Gazette 5(2): 41

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.

Genetic Studies of Mutants that Reiterate Cell Lineages: lin-4 II and unc-86 III

V. Ambros, B. Horvitz

The mutants lin-4(e912) II and unc-86(e1416, e1507, n306) III 
display cell lineage reiterations, in which certain cells appear to 
adopt the lineage potential of one of their ancestors (Chalfie, 
Horvitz, and Sulston, Cold Spring Harbor Abstracts, p.  6, 1979).  As 
a result, some cell types are overproduced and others are not 
generated at all.  We hope to determine how lin-4 and unc-86 control 
cell lineages.  As a first step, we plan to identify the null 
phenotypes of these genes and to establish the nature of the existing 
mutations.  Toward this end, we have begun genetic studies of lin-4 
and unc-86.The three existing alleles of unc-86 III result in similar 
phenotypes: mutant animals are mechanosensory defective (Mec), egg-
laying defective, and Him (producing about 2.5% males).  Surprisingly, 
by both complementation tests and map position, the him defect of 
e1507 is distinct from that of e1416 and n306, i.e.  heterozygotes of 
e1416 or n306 with e1507 are non-Him, and the him defect of e1507 has 
been separated by recombination from the other traits of the original 
e1507 isolate, CB1507.  Perhaps the Him phenotype of CB1507 was 
fortuitous.  None of the three unc-86 alleles is suppressed by the 
null allele-specific suppressor sup-7(st5) X.  We have sought 
revertants of e1416: following EMS mutagenesis, 15,000 F1's and their 
progeny were screened for increased motility (unc-86 animals, like 
other Mec mutants, are lethargic).  An extragenic suppressor was 
isolated and mapped to LGI.  The suppressed animals were active but 
still Mec, and the isolated suppressor appeared to be hyperactive; we 
suspect that this mutant bypasses the Mec-induced lethargy.  We have 
obtained a deficiency by mutagenizing males with gamma-rays, mating 
them with appropriately marked unc-86 hermaphrodites, and seeking Unc-
86 Fl cross progeny.  This deficiency fails to complement for the Mec 
phenotype of CB1507 and for both the Mec and Him phenotypes of e1416; 
it also fails to complement unc-36(e251) and at least two lethal 
mutations we have generated that are closely linked to unc-86.The only 
allele of lin-4 is e912, which was isolated by Babu after P-32 
mutagenesis.  Two independent partial revertants of e912 have been 
obtained; one (n179) appeared spontaneously and the other (n360) was 
isolated after gamma-ray mutagenesis.  n179 and n360 are alleles and 
define a new locus about 4% to the right of dpy-7 X.  The isolated 
suppressor is itself a cell lineage mutant, affecting many of the same 
lineages as e912 (e.g.  vulva, lateral hypodermis, male tail), 
although not as severely.  e912 complements let-22(mn22) and let-30(
mn30), the two most closely linked recessive lethal mutations isolated 
by Bob Herman.  let-30 is to the left of lin-4, and let-22 is to the 
right of lin-4 and to the left of dpy-10; these loci should help 
identify a deficiency spanning lin-4.  e912 is dauer-defective and 
blocks dauer formation by the dauer constitutive mutation daf-4(e1364).
We hope that reversion of daf-4 will yield additional lin-4 alleles; 
the isolation of dauer larvae from mutagenized populations of lin-4; 
rate suppressors of e912.  A new cell lineage 
mutant, n355, has been isolated after gamma-ray mutagenesis.  n355 
appears phenotypically similar to e912; however, in contrast to e912, 
this new mutation is dominant and unlinked to the cluster of LGII.  We 
have compared the SDS gel protein profiles of whole extracts prepared 
from N2, e912, and n355.  Both e912 and n355 are missing an adult-
specific doublet of about 190,000 daltons; the suppressed strain e912; 
n179 has this doublet.  These proteins might be localized in adult-
specific cells that are missing in e912 and n355; alternatively, adult-
specific gene expression may be defective in cells that are present in 
both adults and younger animals.