A second look at cellular mRNA sequences said to function as internal ribosome entry sites.

Abstract:

:This review takes a second look at a set of mRNAs that purportedly employ an alternative mechanism of initiation when cap-dependent translation is reduced during mitosis or stress conditions. A closer look is necessary because evidence cited in support of the internal initiation hypothesis is often flawed. When putative internal ribosome entry sequences (IRESs) are examined more carefully, they often turn out to harbor cryptic promoters or splice sites. This undermines the dicistronic assay, wherein IRES activity is measured by the ability to support translation of the 3' cistron. Most putative IRESs still have not been checked carefully to determine whether the dicistronic vector produces only the intended dicistronic mRNA. The widespread use of the pRF vector is a major problem because this vector, which has Renilla luciferase as the 5' cistron and firefly luciferase as the 3' cistron, has been found to generate spliced transcripts. RNA transfection assays could theoretically circumvent these problems, but most candidate IRESs score very weakly in that test. The practice of calling even very weak results 'positive' is one of the problems discussed herein. The extremely low efficiency of putative IRESs is inconsistent with their postulated biological roles.'

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Kozak M

doi

10.1093/nar/gki958

keywords:

subject

Has Abstract

pub_date

2005-11-28 00:00:00

pages

6593-602

issue

20

eissn

0305-1048

issn

1362-4962

pii

33/20/6593

journal_volume

33

pub_type

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