Functional analysis of the microtubule-interacting transcriptome

Mol Biol Cell. 2011 Nov;22(22):4312-23. doi: 10.1091/mbc.E11-07-0629. Epub 2011 Sep 21.

Abstract

RNA localization is an important mechanism for achieving precise control of posttranscriptional gene expression. Previously, we demonstrated that a subset of cellular mRNAs copurify with mitotic microtubules in egg extracts of Xenopus laevis. Due to limited genomic sequence information available for X. laevis, we used RNA-seq to comprehensively identify the microtubule-interacting transcriptome of the related frog Xenopus tropicalis. We identified ~450 mRNAs that showed significant enrichment on microtubules (MT-RNAs). In addition, we demonstrated that the MT-RNAs incenp, xrhamm, and tpx2 associate with spindle microtubules in vivo. MT-RNAs are enriched with transcripts associated with cell division, spindle formation, and chromosome function, demonstrating an overrepresentation of genes involved in mitotic regulation. To test whether uncharacterized MT-RNAs have a functional role in mitosis, we performed RNA interference and discovered that several MT-RNAs are required for normal spindle pole organization and γ-tubulin distribution. Together, these data demonstrate that microtubule association is one mechanism for compartmentalizing functionally related mRNAs within the nucleocytoplasmic space of mitotic cells and suggest that MT-RNAs are likely to contribute to spindle-localized mitotic translation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Cycle / genetics
  • Cell Cycle Proteins / genetics
  • Cell Division
  • Chromosomal Proteins, Non-Histone / genetics
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / genetics
  • Microtubules / metabolism*
  • Mitosis
  • Nuclear Proteins / genetics
  • Phosphoproteins / genetics
  • RNA Interference
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism*
  • RNA, Small Interfering
  • Sequence Alignment
  • Sequence Analysis, RNA
  • Spindle Apparatus / genetics
  • Spindle Apparatus / metabolism*
  • Transcriptome*
  • Tubulin
  • Xenopus
  • Xenopus Proteins / genetics

Substances

  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone
  • Hmmr protein, Xenopus
  • INCENP protein, Xenopus
  • Microtubule-Associated Proteins
  • Nuclear Proteins
  • Phosphoproteins
  • RNA, Messenger
  • RNA, Small Interfering
  • TPX2 protein, Xenopus
  • Tubulin
  • Xenopus Proteins