Transcription elongation factor SII interacts with a domain of the large subunit of human RNA polymerase II

Journal Article · Mon Aug 01 04:00:00 EDT 1988 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.8.8.3136· OSTI ID:6737041

Rappaport, J; Cho, K; Saltzman, A; Prenger, J; Golomb, M; Weinmann, R

Genomic sequences for the large subunit of human RNA polymerase II corresponding to a part of the fifth exon were inserted into an expression vector at the carboxy-terminal end of the ..beta..-galactosidase gene. The in-frame construct produced a 125-kilodalton fusion protein, containing approximately 10 kilodaltons of the large subunit of RNA polymerase II and 116 kilodaltons of ..beta..-galactosidase. The purified bacterially produced fusion protein inhibited specific transcription from the adenovirus type 2 major late promoter, while ..beta..-galactosidase had no effect. The effect of the fusion protein was during RNA elongation, not at the level of initiation, resembling the faithfully initiated but incomplete transcripts produced with purified factors in the absence of SII. Similarly, monoclonal antibody 2-7B, which reacts with the RNA polymerase II region represented in the fusion protein, inhibited specific transcription at the level of elongation in a whole-cell extract. Both monoclonal antibody 2-7B and the fusion protein, although unable to inhibit purified RNA polymerase II in a nonspecific transcription assay, selectively blocked the stimulation elicited by transcription elongation factor SII on the activity of the purified enzyme in vitro. This suggests that the fusion protein traps the SII in nonstimulatory interactions and that anitibody 2-7B inhibits SII binding to RNA polymerase II. Thus, this suggests that an SII-binding contact required for specific RNA elongation resides within the fifth exon region of the largest RNA polymerase II subunit.

• Research Organization: The Wistar Institute, Philadelphia, PA (US)
• OSTI ID: 6737041
• Journal Information: Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 8:8; ISSN MCEBD
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ADENOVIRUS
ANIMALS
ANTIBODIES
BIOCHEMICAL REACTION KINETICS
DNA SEQUENCING
ENZYME ACTIVITY
ENZYMES
GALACTOSIDASE
GENE REGULATION
GENE REPRESSORS
GLYCOSYL HYDROLASES
HYDROLASES
IN VITRO
KINETICS
MAMMALS
MAN
MICROORGANISMS
MOLECULAR BIOLOGY
MONOCLONAL ANTIBODIES
NUCLEOPROTEINS
NUCLEOTIDYLTRANSFERASES
O-GLYCOSYL HYDROLASES
ONCOGENIC VIRUSES
ORGANIC COMPOUNDS
PARASITES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PRIMATES
PROTEINS
REACTION KINETICS
RNA POLYMERASES
STRUCTURAL CHEMICAL ANALYSIS
TRANSCRIPTION FACTORS
TRANSFERASES
VERTEBRATES
VIRUSES