Identification and intracellular localization of the unc-22 gene product of Caenorhabditis elegans

Genes Dev. 1988 Jan;2(1):93-105. doi: 10.1101/gad.2.1.93.

Abstract

The unc-22 gene is one of a set of genes identified using classical genetics that affect muscle structure and function in the free-living nematode Caenorhabditis elegans. Since cloning the unc-22 gene by transposon tagging, we have used conventional techniques combined with a set of Tc1 transposon insertion alleles to characterize the gene and its products. The gene extends over more than 20 kb of genomic sequence and produces a transcript of approximately 14 kb. A polyclonal antibody raised against an Escherichia coli beta-galactosidase-unc-22 fusion protein recognizes a polypeptide in nematode extracts that is between 500,000 and 600,000 daltons and labels the muscle A-band in indirect immunofluorescent microscopy. The Tc1-induced alleles have been used at every stage to verify these conclusions. The Tc1 insertions are spread over much of the region that contributes to the mature transcript; in most alleles, Tc1 sequences are incorporated into a composite unc-22-Tc1 transcript. The large protein is either absent or severely reduced in amounts in the mutants. In one case, a truncated polypeptide was also identified. The location of the protein in the A-band, along with earlier genetic data, suggests that the unc-22 product may interact with myosin to regulate its function.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Caenorhabditis / genetics*
  • Cell Compartmentation
  • DNA Transposable Elements
  • Fluorescent Antibody Technique
  • Immunosorbent Techniques
  • Molecular Weight
  • Muscle Proteins / genetics*
  • Muscle Proteins / physiology
  • Muscles / physiology*
  • Muscles / ultrastructure
  • Mutation
  • Myosins / metabolism
  • Phenotype
  • RNA, Messenger / genetics

Substances

  • DNA Transposable Elements
  • Muscle Proteins
  • RNA, Messenger
  • Myosins