Expression Patterns and Levels of All Tubulin Isotypes Analyzed in GFP Knock-In C. elegans Strains

Cell Struct Funct. 2021 Jun 30;46(1):51-64. doi: 10.1247/csf.21022. Epub 2021 May 8.

Abstract

Most organisms have multiple α- and β-tubulin isotypes that likely contribute to the diversity of microtubule (MT) functions. To understand the functional differences of tubulin isotypes in Caenorhabditis elegans, which has nine α-tubulin isotypes and six β-tubulin isotypes, we systematically constructed null mutants and GFP-fusion strains for all tubulin isotypes with the CRISPR/Cas9 system and analyzed their expression patterns and levels in adult hermaphrodites. Four isotypes-α-tubulins TBA-1 and TBA-2 and β-tubulins TBB-1 and TBB-2-were expressed in virtually all tissues, with a distinct tissue-specific spectrum. Other isotypes were expressed in specific tissues or cell types at significantly lower levels than the broadly expressed isotypes. Four isotypes (TBA-5, TBA-6, TBA-9, and TBB-4) were expressed in different subsets of ciliated sensory neurons, and TBB-4 was inefficiently incorporated into mitotic spindle MTs. Taken together, we propose that MTs in C. elegans are mainly composed of four broadly expressed tubulin isotypes and that incorporation of a small amount of tissue-specific isotypes may contribute to tissue-specific MT properties. These newly constructed strains will be useful for further elucidating the distinct roles of tubulin isotypes.Key words: tubulin isotypes, microtubules, C. elegans.

Keywords: C. elegans; microtubules; tubulin isotypes.

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / metabolism
  • Caenorhabditis elegans* / genetics
  • Caenorhabditis elegans* / metabolism
  • Gene Knock-In Techniques
  • Green Fluorescent Proteins* / genetics
  • Green Fluorescent Proteins* / metabolism
  • Microtubules / metabolism
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Tubulin* / genetics
  • Tubulin* / metabolism

Substances

  • Tubulin
  • Green Fluorescent Proteins
  • Protein Isoforms
  • Caenorhabditis elegans Proteins