Molecular Biology Reports

, Volume 27, Issue 3, pp 149–156

Identification of a ∼30S size non-ribosomal Saccharomyces cerevisiae RNA that is rapidly labeled on its 3′ end by ATP or UTP

Authors

  • Krishna Sinha
    • Department of PharmacologyBaylor College of Medicine Houston
  • Ram Reddy
    • Department of PharmacologyBaylor College of Medicine Houston
Article

DOI: 10.1023/A:1007139114689

Cite this article as:
Sinha, K. & Reddy, R. Mol Biol Rep (2000) 27: 149. doi:10.1023/A:1007139114689

Abstract

Cell-free extracts prepared from S. cerevisiae cells were incubated in the presence of [α-32P]-labeled ATP, CTP, GTP or UTP. An RNA larger than ribosomal 25S RNA with an apparent size of approximately 30S was prominently labeled on its 3′ end in the presence of ATP or UTP but not with CTP or GTP. This labeled RNA was not hybrid-selected by cloned yeast ribosomal DNA; in addition, this ∼30S RNA was not cleaved by RNase H in the presence of complementary deoxyribooligonucleotides to rRNA. These two lines of evidence show that this ∼30S RNA is not structurally related to ribosomal RNA gene repeat. The cell-free extracts prepared from yeast cells containing temperature-sensitive poly(A) polymerase adenylated this novel yeast RNA at restrictive temperature with efficiency similar to extracts prepared from wild-type yeast cells. These data show that the enzyme responsible for adenylation of this ∼30S RNA is distinct from mRNA poly(A) polymerase. While the human SRP RNA 3′ adenylating enzyme in the HeLa cell extract adenylated human SRP or Alu RNAs, the yeast adenylating enzyme did not adenylate the human SRP or Alu RNAs in vitro; these data indicate species specificity for this adenylating enzyme.

3′ adenylation3′ uridylationpost-transcriptional labelingyeast 30SRNA

Copyright information

© Kluwer Academic Publishers 2000