Worm Breeder's Gazette 11(3): 29

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Analysis of par-2 cDNA

Diane Levitan and Dan Stinchcomb

Figure 1

We have been cloning the maternal-effect lethal gene par-2.  Embryos 
from a homozygous par-2 mother undergo symmetric and synchronous 
cleavages, a failure to localize P granules properly, a low expression 
(20%) of gut granules, and no morphogenesis (Kemphues et al.  Cell 
52:1988).  The role of the wild type par-2 gene, as well as the other 
par genes, is therefore thought to be in the organization of the 
cytoplasm and the proper partitioning of cytoplasmic components in 
early embryogenesis.  We have previously described the identification 
of a par-2 allele specific polymorphism, which consists of a 3 kb 
deletion in the allele it46.  We have also shown that the DNA 
contained within this deletion identifies a 2.3 kb oocyte specific 
message when used to probe a Northern blot containing RNA isolated 
from fem-2 (sperm minus) and glp-1 (germline minus) animals (1989 CSH 
C.  elegans meeting).  Because this message seemed likely to be the 
par-2 message, we used the same probe to isolate cDNA's from the 
Schauer, Kim, Barstead, and Martin cDNA libraries.  Although none of 
these cDNA's are full length, we are within a few hundred base pairs 
of the complete message.  Hybridization experiments with 5' and 3' 
specific cDNA probes indicate that the it46 polymorphism deletes 
coding sequences found in this cDNA.  These data strongly support the 
notion that this cDNA clone represents the par-2 message.
We have sequenced 2.1 kb of this cDNA clone and searched several 
databases with the predicted protein product--however, no homologous 
proteins were found.  The one notable feature of this sequence is the 
presence of an ATP-binding site of the myosin class.  The figure below 
shows the ATP-binding site consensus sequence, the par-2 putative ATP-
binding site, as well as other members of this class of proteins.  
While we don't yet understand the role this protein plays in early 
development, we are intrigued by the presence of the ATP-binding site 
which is consistent with a role for par-2 in cytoplasmic sorting 
during early cleavage divisions.
[See Figure 1]

Figure 1