Worm Breeder's Gazette 14(4): 22 (October 1, 1996)

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.

Partial rescue and further characterization of him-3.

Monique Zetka, Fritz Mueller

Institute of Zoology University of Fribourg Perolles CH-1700 Fribourg Switzerland

        Meiotic mutations in C. elegans have largely been isolated in
screens for mutations that disrupted the  segregation of the X
chromosomes, resulting in an easily identifiable him phenotype.  Further
characterization of these mutations has suggested that they fall into
two general classes: one of which specifically disrupts the segregation
of the X (and is associated with an X-chromosome specific
recombination-defective phenotype) and another which disrupts the
segregation of all chromosomes (a recombination-defective phenotype has
also been reported for him-6 and him-14).  In him-3(e1256) homozygotes,
crossing over is also reduced on the autosomes and has been shown to
correlate with an increase in the frequency of nondisjunction (K.McKim,
Ph.D Thesis).  On the X chromosome, however, e1256 has little effect on
the frequency of exchange (K. McKim, Ph.D Thesis), making difficult an
explanation for the presence of 12% males amongst the self-fertilization
progeny of mutant hermaphrodites.  e1147 is a weak allele that produces
3.5% males amongst the self-progeny of homozygous, hermaphrodites yet
the frequency of crossing over in e1147 homozygotes is also greatly
reduced:  2.0 m.u. in dpy-11 unc-42 (V) (compared to 2.7 m.u. in
controls) and 12.7 m. u. in dpy-7 unc-3 (X) (compared to 21.5 m.u. in
controls).  These results suggest that the him-3 mutations uncouple the
strict relationship between crossing over and proper segregation and
their effects on crossing over in several other intervals is now being

        To determine the molecular basis of the him-3 phenotype, the
rol-6(su1006) transformation plasmid and cosmids in the region to which
him-3 has been genetically mapped on chromosome IV were co-injected into
e1147 hermaphrodites.  The co-injection of ZK616 and F41H10 resulted in
five stable transgenic lines that partially rescued the him phenotype.
The most significant rescuing activity was observed for swEx56, which
segregated 1.1% males and 48% rollers.  To confirm these results, the
extrachromosomal array sEx125 (constructed by D.Collins), which contains
the cosmid ZK1236 from chromosome III, was crossed into the e1147
background.  This strain gave rise to 5.5% males amongst the self
progeny, indicating that the presence of extrachromosomal DNA in general
was not the explanation for the partial rescue observed with F41H10 and
ZK616.  To narrow down the region responsible for the rescuing activity,
ZK616 and F41H10 were injected independently.  A strain carrying the
ZK616-containing array swEx60 segregated 41% rollers and 1.5% males,
whereas four strains carrying the cosmid F41H10 (swEx61-swEx64)
segregated rollers between 20-53% and males between 3.8-5.0%.  This
indicates that the rescuing activity maps to ZK616 and that the general
presence of cosmids from this region is not responsible for the decrease
in the frequency of males.  Deletion derivatives are now being
constructed to narrow down the region responsible and to identify
potential coding elements.  An unexpected result of these experiments
has been the observation that the phenotype of e1147 homozygotes that
bear arrays containing nonrescuing cosmids is more severe than the
canonical phenotype.  For example, the percentage of males rises from
3.5% in e1147 homozygotes to 5.5% in the presence of sEx125 and to 5.0%
in the presence of swEx64 , suggesting that e1147 is sensitiive to the
level of DNA in the cell.

Thanks to Heinz Tobler for his support, Alan Coulson for the cosmids,
Ann Rose and David Baillie for strains, and Jonathan Hodgkin and Phil
Meneely for helpful comments on the rescue data.