Efficient synthesis, termination and release of RNA polymerase III transcripts in Xenopus extracts depleted of La protein

EMBO J. 1998 Apr 1;17(7):2033-41. doi: 10.1093/emboj/17.7.2033.

Abstract

La proteins are conserved, abundant and predominantly nuclear phosphoproteins which bind to the 3'-U termini of newly synthesized RNA polymerase III transcripts. The human La protein has been implicated in the synthesis, termination and release of such transcripts. Here we examine the potential transcriptional properties of La in Xenopus laevis, using a homologous tRNA gene as template. Immunodepletion of La from cell-free extracts leads to the formation of tRNA precursors lacking 3'-U residues. This shortening can be uncoupled from RNA polymerase III transcription, indicating that it results from nuclease degradation rather than incomplete synthesis. Extracts containing <1% of the normal La protein content synthesize tRNA precursors just as well as complete extracts, with no change in termination efficiency, and the vast majority of these full-length transcripts are not associated with the template or with residual La protein. Hence, Xenopus La seems not to function as an initiation, termination or release factor for RNA polymerase III. Consistent with the recently discovered role of La in yeast tRNA maturation in vivo, recombinant Xenopus La prevents 3'-exonucleolytic degradation of tRNA precursors in vitro. A conserved RNA chaperone function may best explain the abundance of La in eukaryotic nuclei.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autoantigens / physiology*
  • Cell Extracts
  • Cell-Free System
  • DNA / metabolism
  • Humans
  • RNA Polymerase III / metabolism*
  • RNA Precursors / metabolism
  • RNA, Transfer, Phe / biosynthesis*
  • RNA, Transfer, Phe / metabolism
  • Ribonucleoproteins / physiology*
  • Templates, Genetic
  • Transcription, Genetic / physiology*
  • Xenopus laevis

Substances

  • Autoantigens
  • Cell Extracts
  • RNA Precursors
  • RNA, Transfer, Phe
  • Ribonucleoproteins
  • SS-B antigen
  • DNA
  • RNA Polymerase III