Molecular cloning of an essential subunit of RNA polymerase II elongation factor SIII.
A transcription factor designated SIII was recently purified from mammalian cells and shown to regulate the activity of the RNA polymerase II elongation complex. SIII is a heterotrimer composed of approximately 110-, 18-, and 15-kDa polypeptides and is capable of increasing the overall rate of RNA chain elongation by RNA polymerase II by suppressing transient pausing of polymerase at multiple sites on the DNA template. Here we describe the molecular cloning and characterization of a cDNA encoding the functional 15-kDa subunit (p15) of SIII. The p15 cDNA encodes a 112-amino-acid polypeptide with a calculated molecular mass of 12,473 Da and an electrophoretic mobility indistinguishable from that of the natural p15 subunit. When combined with the 110- and 18-kDa SIII subunits, bacterially expressed p15 efficiently replaces the natural p15 subunit in reconstitution of transcriptionally active SIII. A homology search revealed that the amino-terminal half of the SIII p15 subunit shares significant sequence similarity with a portion of the RNA-binding domain of Escherichia coli transcription termination protein rho and with the E. coli NusB protein, suggesting that SIII may be evolutionarily related to proteins involved in the control of transcription elongation in eubacteria.
Garrett
K P
KP
Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, Oklahoma City 73104.
Tan
S
S
Bradsher
J N
JN
Lane
W S
WS
Conaway
J W
JW
Conaway
R C
RC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D012319:RNA Polymerase II / Q000235:genetics*; D051381:Rats; D012234:Rho Factor / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; D012234:Rho Factor; D014157:Transcription Factors; D012319:RNA Polymerase II