The block to transcription elongation at the minute virus of mice attenuator is regulated by cellular elongation factors.
We have previously reported that both in vivo and in vitro, RNA polymerase II pauses or prematurely terminates transcription at a specific attenuation site located 142 to 147 nucleotides downstream from the P4 promoter of minute virus of mice (MVM). In this report, we show that an in vitro block to transcription elongation in HeLa whole-cell extract occurs at elevated KCl concentrations (0.2 to 1.5 M) but not at the standard KCl concentration (50 mM). Briefly initiated transcription complexes, devoid of dissociated elongation factors by passage through a Sephacryl S-1000 column at 0.3 M KCl, were allowed to elongate the briefly initiated nascent RNA, and a block to transcription elongation at the attenuation site was observed independently of the KCl concentration at the time of elongation. Moreover, the block to elongation was overcome by the addition, during elongation, to the column of purified complexes of whole-cell extract from EA cells but not from MVM-infected EA cells or HeLa cells. The general transcription factors IIF and IIX were also shown to alleviate this block to transcription elongation. On the basis of these results, we suggest that the block to elongation at the MVM attenuation site observed late in MVM infection results, at least in part, from the inactivation of the general transcription elongation factors.
Krauskopf
A
A
Department of Molecular Genetics and Virology, Weizmann Institute of Science, Rehoyot, Israel.
Bengal
E
E
Aloni
Y
Y
D000818:Animals; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D006367:HeLa Cells / Q000502:physiology; D006801:Humans; D007700:Kinetics; D051379:Mice; D008915:Minute Virus of Mice / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D013698:Templates, Genetic; D014157:Transcription Factors / Q000502:physiology*; D014158:Transcription, Genetic
D004279:DNA, Viral; D014157:Transcription Factors; D012319:RNA Polymerase II