Identification of in vivo target RNA sequences bound by thymidylate synthase

Edward Chu, Tiziana Cogliati, Sitki M. Copur, Aldo Borre, Donna M. Voeller, Carmen J. Allegra, Shoshana Segal

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

We developed an immunoprecipitation-RNA-random PCR (rPCR) method to isolate cellular RNA sequences that bind to the folate-dependent enzyme thymidylate synthase (TS). Using this approach, nine different cellular RNAs that formed a ribonucleoprotein (RNP) complex with thymidylate synthase (TS) in human colon cancer cells were identified. RNA binding experiments revealed that seven of these RNAs bound TS with relatively high affinity (IC50 values ranging from 1.5 to 6 nM). One of the RNAs was shown to encode the interferon (IFN)-induced 15 kDa protein. Western immunoblot analyses demonstrated that the level of IFN-induced 15 kDa protein was significantly decreased in human colon cancer H630-R10 cells compared with parent H630 cells. While the level of IFN-induced 15 kDa mRNA expression was the same in parent and TS-overexpressing cell lines, the level of IFN-induced 15 kDa RNA bound to TS in the form of a RNP complex was markedly higher in H630-R10 cells relative to parent H630 cells. These studies begin to define a number of cellular target RNA sequences with which TS interacts and suggest that these TS protein-cellular RNA interactions may have a biological role.

Original languageEnglish (US)
Pages (from-to)3222-3228
Number of pages7
JournalNucleic acids research
Volume24
Issue number16
DOIs
StatePublished - 1996
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

Fingerprint Dive into the research topics of 'Identification of in vivo target RNA sequences bound by thymidylate synthase'. Together they form a unique fingerprint.

Cite this