Elements which stimulate gene amplification in mammalian cells : role of recombinogenic sequences/structures and transcriptional activation

HSAG-1 is a 3.4 kb mammalian genomic element which has been shown to stimulate the amplification of the pSV2DHFR expression vector in cis when transfected into a variety of cell lines (1). This amplification stimulatory activity requires the interaction of multiple positive acting elements that incl...

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Bibliographic Details
Published in:Nucleic acids research 1991-05, Vol.19 (9), p.2477-2484
Main Authors: MCARTHUR, J. G, BEITEL, L. K, CHAMBERLAIN, J. W, STANNERS, C. P
Format: Article
Language:English
Subjects:
Animals
Base Composition
Base Sequence
Biological and medical sciences
Cell Line
Cloning, Molecular
Computer Simulation
DNA - genetics
DNA Replication
Fundamental and applied biological sciences. Psychology
Gene Amplification
Gene Expression
Genetic Vectors
Genic rearrangement. Recombination. Transposable element
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Nucleic Acid Conformation
Repetitive Sequences, Nucleic Acid
Restriction Mapping
Transcription, Genetic
Transfection
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Summary:HSAG-1 is a 3.4 kb mammalian genomic element which has been shown to stimulate the amplification of the pSV2DHFR expression vector in cis when transfected into a variety of cell lines (1). This amplification stimulatory activity requires the interaction of multiple positive acting elements that include sequence features associated with recombination 'hotspots', such as Alu-like repetitive sequences and A/T rich regions (2). We demonstrate here that two other members of the HSAG family of elements, HSAG-2 and HSAG-5, also stimulate vector amplification. By analysis of the HSAG-2 nucleotide sequence and of the amplification activity of HSAG-2 and HSAG-5 subfragments, we show that this activity also involves the interaction of multiple positive acting elements. The autonomous replication of the HSAG containing vectors is not responsible for this effect. We also show that the orientation of HSAG elements in pSV2DHFR has a profound effect on their amplification stimulatory activity, and present evidence that the transcription of these elements in pSV2DHFR is necessary for the effect.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/19.9.2477