A dominant negative mutation in a spliceosomal ATPase affects ATP hydrolysis but not binding to the spliceosome. Academic Article uri icon

Overview

abstract

  • PRP16 is an RNA-dependent ATPase required for the second catalytic step of splicing in vitro. A dominant suppressor of a branchpoint mutation in Saccharomyces cerevisiae, the prp16-1 allele, contains a Tyr to Asp change in the nucleotide-binding site consensus sequence. We now find that cells harboring the prp16-1 allele have a general growth defect that is exacerbated at cold temperatures. The mutant is dominant over the wild-type gene when overexpressed. Purified Prp16-1 protein binds to the spliceosome with apparently wild-type affinity; however, it only weakly complements the second-step block in a PRP16-depleted extract. Analysis of purified Prp16-1 revealed that the rate of ATP hydrolysis is greatly reduced. These results can account for the dominant negative growth phenotype and argue that the ATPase activity of PRP16 is essential for its role in splicing. Moreover, since PRP16 is a member of the DEAD/H box families, these findings have important implications for a large class of proteins.

publication date

  • August 1, 1992

Research

keywords

  • Adenosine Triphosphatases
  • Fungal Proteins
  • Mutation
  • RNA Splicing
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins

Identity

PubMed Central ID

  • PMC364619

Scopus Document Identifier

  • 0026656540

Digital Object Identifier (DOI)

  • 10.1128/mcb.12.8.3540-3547.1992

PubMed ID

  • 1385854

Additional Document Info

volume

  • 12

issue

  • 8