Abstract
The eukaryotic transcription factor TFIIS enhances elongation and nascent transcript cleavage activities of RNA polymerase II in a stalled elongation complex. By site-directed mutagenesis, we have demonstrated that invariant residues Asp-261 and Glu-262 of the nucleic acid-binding TFIIS Zn ribbon are critical for stimulation of both elongation and RNA cleavage activities of RNA polymerase II. Substitution of either of these residues inactivates both TFIIS functions, suggesting a related role in both activities. These acidic residues may participate in phosphoryl transfer reactions by a two-metal-ion mechanism in a manner analogous to Klenow fragment. The RNA polymerase II itself may contain a Zn ribbon, in as much as the polymerase's 15-kDa subunit contains a sequence that aligns well with the TFIIS Zn ribbon sequence, including a similarly placed pair of acidic residues.
MeSH Terms
Amino Acid Sequence
Animals
Base Sequence
Cattle
In Vitro Techniques
Molecular Sequence Data
Mutagenesis, Site-Directed
RNA Polymerase II/metabolism
RNA Processing, Post-Transcriptional
RNA, Messenger/metabolism
Sequence Alignment
Sequence Homology, Amino Acid
Structure-Activity Relationship
Transcription Factors/chemistry,metabolism
Transcription Factors, General
Transcription, Genetic
Transcriptional Elongation Factors
Zinc
Chemicals
RNA, Messenger
Transcription Factors
Transcription Factors, General
Transcriptional Elongation Factors
transcription factor S-II
RNA Polymerase II
Zinc
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Jeon C
Department of Biochemistry, University of Chicago, IL 60637.
Yoon H
Agarwal K
References (23)
23 references, click to expand
-
DNA-dependent RNA polymerase from Ehrlich ascites tumor cells. 3. Ribonuclease H and elongating activity of stimulatory factor S-II.
J Biochem. 1973 Dec;74(6):1177-82
PMID: 4798604
-
RNA polymerase II transcription terminates at a specific DNA sequence in a HeLa cell-free reaction.
Proc Natl Acad Sci U S A. 1986 Oct;83(20):7623-7
PMID: 2429312
-
Rapid and efficient site-specific mutagenesis without phenotypic selection.
Methods Enzymol. 1987;154:367-82
PMID: 3323813
-
Sequence requirements for premature termination of transcription in the human c-myc gene.
Cell. 1988 Apr 22;53(2):245-56
PMID: 2834065
-
Isolation of a recombinant copy of the gene encoding C/EBP.
Genes Dev. 1988 Jul;2(7):786-800
PMID: 2850264
-
Transcription elongation factor SII (TFIIS) enables RNA polymerase II to elongate through a block to transcription in a human gene in vitro.
J Biol Chem. 1989 Jun 25;264(18):10799-809
PMID: 2471707
-
Adenosine deaminase gene expression. Tissue-dependent regulation of transcriptional elongation.
J Biol Chem. 1989 Aug 25;264(24):14561-5
PMID: 2474547
-
RNA polymerase II elongation complexes paused after the synthesis of 15- or 35-base transcripts have different structures.
Mol Cell Biol. 1991 Mar;11(3):1508-22
PMID: 1705007
-
Cloning, expression and characterization of the human transcription elongation factor, TFIIS.
Nucleic Acids Res. 1991 Mar 11;19(5):1073-9
PMID: 1708494
-
Stimulation of transcript elongation requires both the zinc finger and RNA polymerase II binding domains of human TFIIS.
Biochemistry. 1991 Aug 6;30(31):7842-51
PMID: 1868060
-
Yeast RNA polymerase II subunit RPB9 is essential for growth at temperature extremes.
J Biol Chem. 1991 Oct 5;266(28):19053-5
PMID: 1918023
-
Purification of his-tagged proteins in non-denaturing conditions suggests a convenient method for protein interaction studies.
Nucleic Acids Res. 1991 Nov 25;19(22):6337-8
PMID: 1956801
-
Elongation factor-dependent transcript shortening by template-engaged RNA polymerase II.
J Biol Chem. 1992 Feb 25;267(6):3795-800
PMID: 1371280
-
The elongation-termination decision in transcription.
Science. 1992 Feb 14;255(5046):809-12
PMID: 1536005
-
The RNA polymerase II 15-kilodalton subunit is essential for viability in Drosophila melanogaster.
Mol Cell Biol. 1992 Mar;12(3):928-35
PMID: 1545824
-
Netropsin specifically enhances RNA polymerase II termination at terminator sites in vitro.
Proc Natl Acad Sci U S A. 1992 May 1;89(9):3676-80
PMID: 1315032
-
Crystal structure at 3.5 A resolution of HIV-1 reverse transcriptase complexed with an inhibitor.
Science. 1992 Jun 26;256(5065):1783-90
PMID: 1377403
-
The RNA polymerase II ternary complex cleaves the nascent transcript in a 3'----5' direction in the presence of elongation factor SII.
Genes Dev. 1992 Jul;6(7):1342-56
PMID: 1378419
-
GreA protein: a transcription elongation factor from Escherichia coli.
Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8899-902
PMID: 1384037
-
Identification of a 3'-->5' exonuclease activity associated with human RNA polymerase II.
Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):843-7
PMID: 8381534
-
Structure of a new nucleic-acid-binding motif in eukaryotic transcriptional elongation factor TFIIS.
Nature. 1993 Sep 16;365(6443):277-9
PMID: 7626141
-
Novel zinc finger motif in the basal transcriptional machinery: three-dimensional NMR studies of the nucleic acid binding domain of transcriptional elongation factor TFIIS.
Biochemistry. 1993 Sep 28;32(38):9944-59
PMID: 8399164
-
RNA polymerase II transcription factor SIII. I. Identification, purification, and properties.
J Biol Chem. 1993 Dec 5;268(34):25587-93
PMID: 8244996