Worm Breeder's Gazette 11(2): 72

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.

Cellular Proteins Bind to the Nematode Trans-spliced RNA Leader Sequence

Xin-Yun Huang and David Hirsh

The trans-spliced RNA leader might play a role in regulating the 
translation and/or stability of SL-containing mRNAs.  This possibility 
led us to seek transacting factors that might mediate this regulatory 
function.  Protein factors from C.  elegans cells (either embryos or 
worms from mixed stages) that bind with high specificity to the trans-
spliced RNA leader sequence (SL) have been identified.  Whole cell 
extracts were shown by a mobility shift electrophoresis assay to form 
two RNA-protein complexes involving the SL1 sequence.  Complex 
formation could be abolished by preincubation with excess, unlabeled 
competitor SL1-containing RNA (which is the 5' non-coding region of 
actin-1 mRNA fused to the firefly luciferase coding region) but not by 
nonspecific, control RNA that did not contain an SL1 sequence.
Spliced leader binding protein-1 (SLBP1) is distinct from SLBP2.  
The binding of these proteins to SL1 is cap dependent though 
independent of the detailed structure of the cap; either monomethyl- 
or trimethyl guanosine cap stimulates binding.
We are purifying SLBP1 and SLBP2.  Future work will be focused on 
the characterization of these proteins and elucidation of their 
biological functions.