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PMID: 8265584 Published · ppublish English Journal Article

Translation of the human LINE-1 element, L1Hs.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 90 ·No. 24 ·1993-12-15 ·Pages 11533-7

McMillan JP, Singer MF

Abstract

Full-length RNA transcribed from the human LINE-1 (L1) element L1 Homo sapiens (L1Hs) has a 900-nt, G+C-rich, 5'-untranslated region (UTR). The 5' UTR is followed by two long open reading frames, ORF1 and ORF2, which are separated from each other by an inter-ORF region of 33 nt that includes two or three in-frame stop codons. We examine here the mechanism(s) by which the translation of L1Hs ORF1 and ORF2 is initiated. A stable hairpin structure (delta G = -74.8 kcal/mol), inserted at nt 661 of the 5' UTR, caused a 3- to 8-fold decrease in the in vitro and in vivo translation of either a lacZ reporter gene for ORF1 or the ORF1 polypeptide product, p40, but translation of a lacZ reporter gene in ORF2 was increased. The results are compatible with a model for ORF1 translation initiation in which the majority of ribosomes scan from a point 5' of nt 661 but suggest that ORF2 is not translated by attached ribosomes that reinitiate after the termination of ORF1 translation. Our data are compatible with a model whereby the translation of L1Hs ORF2 is initiated internally.

MeSH Terms
Animals Base Sequence Blotting, Northern Calorimetry Cell Line Codon/genetics Genes Hominidae/genetics Humans Molecular Sequence Data Mutagenesis, Insertional Nucleic Acid Conformation Oligodeoxyribonucleotides Open Reading Frames Protein Biosynthesis RNA, Messenger/metabolism Repetitive Sequences, Nucleic Acid Restriction Mapping Transcription, Genetic Transfection beta-Galactosidase/biosynthesis
Chemicals
Codon Oligodeoxyribonucleotides RNA, Messenger beta-Galactosidase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
McMillan J P
Laboratory of Biochemistry, National Cancer Institute, Bethesda, MD 20892.
Singer M F
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.90.24.11533
Published
1993-12-15
Pages
11533-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC48018
Subset
IM
Databases
GENBANK
M80343
External Links
PubMed source DOI: 10.1073/pnas.90.24.11533 PMC Full Text View journal details
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