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In vitro premature termination in SV40 late transcription.
Author(s) -
Pfeiffer P.,
Hay N.,
Pruzan R.,
Jakobovits E.B.,
Aloni Y.
Publication year - 1983
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1983.tb01403.x
Subject(s) - biology , transcription (linguistics) , in vitro , microbiology and biotechnology , genetics , philosophy , linguistics
Nuclear extracts and viral transcribing minichromosomes were prepared from SV40‐infected cells and incubated in vitro with [alpha‐32P]UTP under conditions which allow the elongation of preinitiated RNA chains. Sucrose gradient lysis of the transcription mixtures revealed two populations of SV40‐specific RNA: elongating chains that remain associated with the viral minichromosomes, and, at the top of the gradient, small free RNA detached from the template and hybridizing exclusively to the promoter‐proximal region of SV40 DNA. This free RNA was shown by polyacrylamide gel electrophoresis to comprise essentially a 94 nucleotide species, which could, however, at high UTP concentration, be elongated a further few nucleotides before terminating. These results thus show that the actively transcribing minichromosomes provide a sytem in which the attenuated RNA can be released from the template. Moreover, this is the first demonstration of specific in vitro termination of polymerase B transcription. The conditions which lead to transcription termination are discussed.