scholarly journals Single molecule studies of RNA polymerase II transcription in vitro

Transcription ◽  
2014 ◽  
Vol 5 (1) ◽  
pp. e27608 ◽  
Author(s):  
Abigail E Horn ◽  
James A Goodrich ◽  
Jennifer F Kugel
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Single Molecule ◽  
Single Molecule Studies ◽  
Transcription In Vitro ◽  
Rna Polymerase Ii Transcription

[38] RNA polymerase II transcription in vitro

1991 ◽  
pp. 545-550 ◽  
Author(s):  
Neal F. Lue ◽  
Peter M. Flanagan ◽  
Raymond J. Kelleher ◽  
Aled M. Edwards ◽  
Roger D. Kornberg
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Transcription In Vitro ◽  
Rna Polymerase Ii Transcription

scholarly journals Heterogeneous nuclear ribonucleoprotein K is a transcription factor.

1996 ◽  
Vol 16 (5) ◽  
pp. 2350-2360 ◽  
Author(s):  
E F Michelotti ◽  
G A Michelotti ◽  
A I Aronsohn ◽  
D Levens
Keyword(s):  
Transcription Factor ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Hnrnp K ◽  
Heterogeneous Nuclear Ribonucleoprotein ◽  
Transcription In Vitro ◽  
Nuclear Ribonucleoprotein ◽  
Rna Polymerase Ii Transcription

The CT element is a positively acting homopyrimidine tract upstream of the c-myc gene to which the well-characterized transcription factor Spl and heterogeneous nuclear ribonucleoprotein (hnRNP) K, a less well-characterized protein associated with hnRNP complexes, have previously been shown to bind. The present work demonstrates that both of these molecules contribute to CT element-activated transcription in vitro. The pyrimidine-rich strand of the CT element both bound to hnRNP K and competitively inhibited transcription in vitro, suggesting a role for hnRNP K in activating transcription through this single-stranded sequence. Direct addition of recombinant hnRNP K to reaction mixtures programmed with templates bearing single-stranded CT elements increased specific RNA synthesis. If hnRNP K is a transcription factor, then interactions with the RNA polymerase II transcription apparatus are predicted. Affinity columns charged with recombinant hnRNP K specifically bind a component(s) necessary for transcription activation. The depleted factors were biochemically complemented by a crude TFIID phosphocellulose fraction, indicating that hnRNP K might interact with the TATA-binding protein (TBP)-TBP-associated factor complex. Coimmunoprecipitation of a complex formed in vivo between hnRNP K and epitope-tagged TBP as well as binding in vitro between recombinant proteins demonstrated a protein-protein interaction between TBP and hnRNP K. Furthermore, when the two proteins were overexpressed in vivo, transcription from a CT element-dependent reporter was synergistically activated. These data indicate that hnRNP K binds to a specific cis element, interacts with the RNA polymerase II transcription machinery, and stimulates transcription and thus has all of the properties of a transcription factor.

scholarly journals Uncoupling of initiation and reinitiation rates during HeLa RNA polymerase II transcription in vitro.

1993 ◽  
Vol 13 (8) ◽  
pp. 4572-4577 ◽  
Author(s):  
Y Jiang ◽  
J D Gralla
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Promoter Activity ◽  
Atp Hydrolysis ◽  
Preinitiation Complex ◽  
Transcription In Vitro ◽  
Rna Polymerase Ii Transcription

RNA polymerase II transcription is influenced both by how rapidly a gene is induced and by the rate at which continuous reinitiation occurs after induction. We show here that in vitro the rates of these two critical steps need not be the same. For activator GAL-AH-dependent HeLa transcription, the rate of assembling a preinitiation complex is significantly slower than the rate of reinitiation. Although reinitiation is rapid, it still requires ATP hydrolysis. This unexpected uncoupling of the rates of initiation and reinitiation implies that in regulating mammalian promoter activity, one must consider separately the controls on initiation during induction and the controls on the subsequent reinitiation events.

scholarly journals Single-Molecule Studies of Human RNA Polymerase Ii Transcription Initiation: Core-Promoter Recognition

2011 ◽  
Vol 100 (3) ◽  
pp. 65a
Author(s):  
Alexandros Pertsinidis ◽  
Sang Ryul Park ◽  
Robert Coleman ◽  
Andrei Revyakin ◽  
Robert Tjian ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Single Molecule ◽  
Transcription Initiation ◽  
Core Promoter ◽  
Promoter Recognition ◽  
Single Molecule Studies ◽  
Rna Polymerase Ii Transcription

Uncoupling of initiation and reinitiation rates during HeLa RNA polymerase II transcription in vitro

1993 ◽  
Vol 13 (8) ◽  
pp. 4572-4577
Author(s):  
Y Jiang ◽  
J D Gralla
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Promoter Activity ◽  
Atp Hydrolysis ◽  
Preinitiation Complex ◽  
Transcription In Vitro ◽  
Rna Polymerase Ii Transcription

RNA polymerase II transcription is influenced both by how rapidly a gene is induced and by the rate at which continuous reinitiation occurs after induction. We show here that in vitro the rates of these two critical steps need not be the same. For activator GAL-AH-dependent HeLa transcription, the rate of assembling a preinitiation complex is significantly slower than the rate of reinitiation. Although reinitiation is rapid, it still requires ATP hydrolysis. This unexpected uncoupling of the rates of initiation and reinitiation implies that in regulating mammalian promoter activity, one must consider separately the controls on initiation during induction and the controls on the subsequent reinitiation events.

A mediator required for activation of RNA polymerase II transcription in vitro

Nature ◽  
1991 ◽  
Vol 350 (6317) ◽  
pp. 436-438 ◽  
Author(s):  
Peter M. Flanagan ◽  
Raymond J. Kelleher ◽  
Michael H. Sayre ◽  
Herbert Tschochner ◽  
Roger D. Kornberg
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Transcription In Vitro ◽  
Rna Polymerase Ii Transcription
Sign in / Sign up

Export Citation Format

Share Document