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PMID: 6095042 Published · ppublish English Journal Article

The long terminal repeat of an endogenous intracisternal A-particle gene functions as a promoter when introduced into eucaryotic cells by transfection.

Molecular and cellular biology ·Vol. 4 ·No. 10 ·1984-10-00 ·Pages 2128-35

Lueders KK, Fewell JW, Kuff EL, Koch T

Abstract

We describe experiments designed to determine whether an endogenous intracisternal A-particle (IAP) gene randomly selected from a mouse embryo library has the potential to be transcriptionally active. Assays for IAP gene transcription were done with permanently transformed rat cells and transiently transfected monkey and mouse cells. The rat cells, which had integrated IAP gene copies, contained IAP RNA. A start site within the IAP 5' long terminal repeat (LTR) was localized by S1 mapping. The promoter activity of the IAP LTR was also measured in cells 48 h after the introduction of recombinant plasmids in which bacterial chloramphenicol acetyl transferase (CAT) encoding sequences were under the control of the LTR. The IAP LTR promoted CAT activity in mouse and monkey cells. In mouse L-cells, the levels of CAT activity were 10 to 25% of those promoted by an analogous recombinant containing the Moloney murine sarcoma virus LTR as the promoter. In contrast to the Moloney murine sarcoma virus LTR, the IAP LTR was five- to eightfold more active in monkey cells than in mouse cells. The 5' and 3' LTRs were equally active, and promoter activity was dependent on having the orientation of the LTRs with respect to the CAT gene the same as their orientation with respect to the IAP gene. A 5'-flanking sequence containing a member of the highly repetitive R-sequence family increased CAT activity in COS cells 11-fold when present along with the LTR. Our results indicate that the LTR of an endogenous mouse IAP gene can function as an efficient promoter in heterologous as well as homologous cells.

MeSH Terms
Acetyltransferases/metabolism Animals Base Sequence Cell Line Chloramphenicol O-Acetyltransferase Cloning, Molecular Endonucleases/metabolism Gene Expression Regulation Mice Operon Plasmids Repetitive Sequences, Nucleic Acid Retroviridae/genetics Single-Strand Specific DNA and RNA Endonucleases Transcription, Genetic Transfection
Chemicals
Acetyltransferases Chloramphenicol O-Acetyltransferase Endonucleases Single-Strand Specific DNA and RNA Endonucleases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lueders K K
Fewell J W
Kuff E L
Koch T
References (49)
49 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
Published
1984-10-00
Pages
2128-35
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC369031
Subset
IM
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