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PMID: 1380649 Published · ppublish English Journal Article

Translation of the rat LINE bicistronic RNAs in vitro involves ribosomal reinitiation instead of frameshifting.

Molecular and cellular biology ·Vol. 12 ·No. 9 ·1992-09-00 ·Pages 4242-8

Ilves H, Kahre O, Speek M

Abstract

The genomic structure of the rat LINE (L1Rn) DNA element contains two overlapping open reading frames (ORFs) and apparently has a potential to code for a DNA/RNA-binding protein (in ORF1) and a reverse transcriptase (in ORF2). We have characterized a 1,630-bp L1Rn cDNA clone encompassing the overlapping ORFs and a 600-bp genomic fragment derived from a full-length L1Rn member and containing the beginning of ORF1. These DNAs were used to restore in part the ORF1-ORF2 organization of L1Rn after being cloned into the pSP65 vector under the control of SP6 polymerase promoter. To test whether L1Rn ORF1 and ORF2 are expressed as a fusion protein, a series of capped RNAs with progressive truncations containing one or both ORFs were prepared and translated in the rabbit reticulocyte lysate. Our analysis indicates that the expression of a putative reverse transcriptase-encoded L1Rn ORF2 in vitro is regulated by reinitiation or internal initiation of translation but not by ribosomal frameshifting.

MeSH Terms
Amino Acid Sequence Animals Base Sequence DNA DNA Transposable Elements DNA-Binding Proteins/genetics Molecular Sequence Data Open Reading Frames/genetics Protein Biosynthesis RNA, Messenger/genetics,metabolism RNA-Binding Proteins/genetics RNA-Directed DNA Polymerase/genetics Rats Repetitive Sequences, Nucleic Acid Ribosomes/metabolism
Chemicals
DNA Transposable Elements DNA-Binding Proteins RNA, Messenger RNA-Binding Proteins DNA RNA-Directed DNA Polymerase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ilves H
Laboratory of Molecular Genetics, Institute of Chemical Physics and Biophysics, Tartu, Estonia.
Kahre O
Speek M
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36 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.9.4242-4248.1992
Published
1992-09-00
Pages
4242-8
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360333
Subset
IM
Databases
GENBANK
X61294, X61295, X61296, X61297, X61298
External Links
PubMed source DOI: 10.1128/mcb.12.9.4242-4248.1992 PMC Full Text View journal details
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