Ordered assembly of a functional preinitiation transcription complex, containing vaccinia virus early transcription factor and RNA polymerase, on an immobilized template.
A functional preinitiation transcription complex was formed by incubating vaccinia virus early transcription factor VETF and RAP94+ RNA polymerase with an early promoter template immobilized on paramagnetic particles. A preferred order of assembly, VETF followed by RNA polymerase, was demonstrated by stepwise addition experiments. ATP was unnecessary for preinitiation transcription complex formation, but divalent cations were required specifically for the association of RNA polymerase.
Baldick
C J
CJ
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
Cassetti
M C
MC
Harris
N
N
Moss
B
B
D001483:Base Sequence; D002413:Cations, Divalent; D004268:DNA-Binding Proteins / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D013698:Templates, Genetic; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014616:Vaccinia virus / Q000235:genetics*; D014764:Viral Proteins / Q000378:metabolism*
D002413:Cations, Divalent; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D014157:Transcription Factors; D014764:Viral Proteins; C064449:early transcription factor protein, Vaccinia virus; D012321:DNA-Directed RNA Polymerases