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Resources » Paper

Aronoff R et al. (1996) West Coast Worm Meeting "PLASMID RESCUE FINALLY. IS SMG-4 A PROTEIN WITH AN RRM AND A KE RICH DOMAIN?"

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  • Comments on Aronoff R et al. (1996) West Coast Worm Meeting "PLASMID RESCUE FINALLY. IS SMG-4 A PROTEIN WITH AN RRM AND A KE RICH DOMAIN?" (0)

  • Overview

    Status:
    Publication type:
    Meeting_abstract
    WormBase ID:
    WBPaper00017298

    Aronoff R, Baran R, & Hodgkin JA (1996). PLASMID RESCUE FINALLY. IS SMG-4 A PROTEIN WITH AN RRM AND A KE RICH DOMAIN? presented in West Coast Worm Meeting. Unpublished information; cite only with author permission.

    The seven smg genes mediate mRNA surveillance, a system that detects and destroys aberrant mRNA, in the nematode C. elegans. Previously cloned smg genes encode an RNA helicase, a protein kinase and several novel products, some of which contain motifs implicated in protein:protein interactions and can be shown to co-immunoprecipitate in vitro (Anderson lab, unpub'd results). Rescue of smg-4 mutants, assayed in an unc-54(r293);smg-4 (ma116) strain, was previously obtained with yeast artificial chromosomes, Y1D1 and Y42F5. Phage were isolated from a library generated from Y42F5 genomic DNA, and one clone, gmRB162, was found to rescue smg-4 mutant animals. Plasmid subclones were tested in further rescue assays, and the predicted gene sequence analyzed. The smallest rescuing subclone of 3.6Kbp, pgm185, could encode two potential open reading frames, which may be part of an operon. Subclones containing either ORF will not rescue. However, when only one or the other ORF is disrupted by site directed mutagenesis, only mutation of the downstream ORF will abrogate rescuing activity. So it seems the rescuing activity requires upstream sequences for its expression, and the downstream ORF mediates smg-4+ function. A 1.5Kb RNA is detected in both total and pA+ RNA, which could encode either predicted gene product. Restriction fragment length polymorphisms have been found associated with mutant smg-4 alleles by southern blotting. For instance, 2.5Kbp is deleted from this region in smg-4 (r1169), an allele generated with DEO by S. Kuchma. This 2.5Kb includes a domain which contains a good match to the octamer consensus sequence of the RNA Recognition Motif (RRM), an 80 a.a. RNA binding motif. In addition, the ORF has a lysine and glutamic acid (KE) rich domain which shares sequence identity with a few genes, of particular note, the yeast gene SPB. Analysis of cDNAs, sequencing of mutant alleles, and expression and more rescue studies are currently in progress.

    Affiliation:
    - Center for the Molecular Biology of RNA, UC Santa Cruz, CA 95064 Department of Molecular and Cell Biology, UC Berkeley, CA 94720 MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH ENGLAND


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