1997
DOI: 10.1074/jbc.272.41.25851
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RNA Polymerase II Subunits 2, 3, and 11 Form a Core Subassembly with DNA Binding Activity

Abstract: RNA polymerase II purified from the fission yeast Schizosaccharomyces pombe consists of 10 species of subunit polypeptide. We introduced a histidine cluster tag sequence into the chromosomal rpb1 and rpb3 genes, which encode subunit 1 (Rpb1) and subunit 3 (Rpb3), respectively, and purified the RNA polymerase by Ni 2؉ affinity chromatography. After stepwise dissociation of the Rpb1-and Rpb3-tagged RNA polymerases fixed on Ni 2؉ -resin by increasing concentrations of urea or guanidium hydrochloride, Rpb2-Rpb3-Rp… Show more

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Cited by 71 publications
(98 citation statements)
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“…One plausible role for the [4Fe-4S] clusters is to regulate the assembly of RNAP in response to cellular conditions, and this is supported by previous studies documenting archaeal subunit D, bacterial ␣ subunit, and eukaryotic Rpb3 as key subunits that initiate the assembly of RNAP (9,10,37,38). The D/␣/Rpb3 subunits are not directly involved in binding of template DNA and synthesis of RNA.…”
Section: Fe-4s] Clusters Of M Acetivorans Subunit D Are Oxygenlabilsupporting
confidence: 59%
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“…One plausible role for the [4Fe-4S] clusters is to regulate the assembly of RNAP in response to cellular conditions, and this is supported by previous studies documenting archaeal subunit D, bacterial ␣ subunit, and eukaryotic Rpb3 as key subunits that initiate the assembly of RNAP (9,10,37,38). The D/␣/Rpb3 subunits are not directly involved in binding of template DNA and synthesis of RNA.…”
Section: Fe-4s] Clusters Of M Acetivorans Subunit D Are Oxygenlabilsupporting
confidence: 59%
“…The [4Fe-4S] clusters in RNAP may serve a similar role. Based on previous structure-function studies of bacterial ␣ N-terminal domain (36,37), eukaryotic Rpb3 (32,38), and archaeal D of RNAPs (2) Fig. 10.…”
Section: Fe-4s] Clusters Of M Acetivorans Subunit D Are Oxygenlabilmentioning
confidence: 99%
“…The enzyme is composed of 12 subunits, Rpb1 to Rpb12, similar to what has been described for the budding yeast S. cerevisiae and mammalian cells (2,3). Biochemical analysis of the enzyme has revealed the presence of several distinct subcomplexes and has proved important in the analysis of the recently published x-ray crystallography structure of S. cerevisiae pol II (4,5). Previous characterization of S. pombe TFIIH has only concerned a specific subcomplex of this transcription factor, containing the Mcs6, Mcs2, and Pmh1 gene products (6).…”
mentioning
confidence: 82%
“…coli RNAP assembles in vivo and in vitro according to the following scheme: ␣-␣ 2 -␣ 2 ␤-␣ 2 ␤␤Ј (9). The ␣ 2 ␤ assembly intermediate appears to be evolutionary conserved, and an ␣ 2 ␤-like RNAP II subassembly was isolated from yeasts (10,11). Further, mutations in E. coli ␣, and in its yeast RNAP II counterpart, Rpb3, that affect the ␣ 2 ␤ formation in their respective systems occur in homologous positions (12,13).…”
mentioning
confidence: 99%