Transcription elongation factor SII

Wind, Megan; Reines, Daniel

doi:10.1002/(sici)1521-1878(200004)22:4<327::aid-bies3>3.0.co;2-4

Cited by 187 publications

(169 citation statements)

References 93 publications

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“…In the presence of the cleavage factor TFIIS, templated GMP incorporation with scaffold A occurred normally, but nontemplated AMP incorporation was suppressed (Fig. 2g), consistent with a role of TFIIS in transcriptional proofreading 16,17 . However, TFIIS was unable to promote passage over the translocation barrier.…”

Section: Impaired Entry Of Lesions Into the Active Sitesupporting

confidence: 62%

Mechanism of transcriptional stalling at cisplatin-damaged DNA

Damsma

Alt

Brueckner

et al. 2007

Nat Struct Mol Biol

160

133

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The anticancer drug cisplatin forms 1,2-d(GpG) DNA intrastrand cross-links (cisplatin lesions) that stall RNA polymerase II (Pol II) and trigger transcription-coupled DNA repair. Here we present a structure-function analysis of Pol II stalling at a cisplatin lesion in the DNA template. Pol II stalling results from a translocation barrier that prevents delivery of the lesion to the active site. AMP misincorporation occurs at the barrier and also at an abasic site, suggesting that it arises from nontemplated synthesis according to an 'A-rule' known for DNA polymerases. Pol II can bypass a cisplatin lesion that is artificially placed beyond the translocation barrier, even in the presence of a G . A mismatch. Thus, the barrier prevents transcriptional mutagenesis. The stalling mechanism differs from that of Pol II stalling at a photolesion, which involves delivery of the lesion to the active site and lesion-templated misincorporation that blocks transcription.Cisplatin (cis-diamminedichloroplatinum(II)) is a widely used anticancer drug that forms DNA adducts that interfere with replication and transcription 1 . The most frequent cisplatin DNA adducts are 1,2-d(GpG) intrastrand cross-links, or cisplatin lesions, in which platinum coordinates the N7 atoms of adjacent guanosines in a DNA strand 2 (Fig. 1a). A cisplatin lesion in the DNA template strand blocks transcription elongation by the single-subunit RNA polymerase from phage T7 (ref.3) and by Pol II 4-6 , and leads to stable polymerase stalling 7 . The stalled Pol II elongation complex can be bound by the elongation factor TFIIS, which stimulates polymerase back-tracking and 3¢ RNA cleavage 6 . A small fraction of polymerases can apparently read through a cisplatin lesion 6 .The detailed molecular mechanisms of cisplatin DNA adduct processing by nucleic acid polymerases are not understood. Here we have used a combination of X-ray crystallography and RNA-extension assays to derive the molecular mechanism of Saccharomyces cerevisiae Pol II stalling at a cisplatin lesion. Comparison of the results with our previous analysis of Pol II stalling at a DNA photolesion, a TT cyclobutane pyrimidine dimer 8 (CPD, Fig. 1a), reveals that the two types of dinucleotide lesions trigger transcriptional stalling by different mechanisms. RESULTS Structure of a cisplatin-damaged Pol II elongation complexTo elucidate recognition of cisplatin-induced DNA damage by transcribing Pol II, we carried out a structure-function analysis of elongation complexes containing a cisplatin lesion in the template DNA strand. Elongation complexes were reconstituted from the 12-subunit S. cerevisiae Pol II and nucleic acid scaffolds as described 8,9 .A cisplatin lesion was first incorporated at registers +2/+3 of the template strand, directly downstream of the NTP-binding site at register +1 (scaffold A, Fig. 1b). The crystal structure of the resulting cisplatin-damaged elongation complex (complex A) was determined at 3.8-Å resolution (Fig. 1c,d and Methods). A very strong peak in the anomalous differe...

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Section: Impaired Entry Of Lesions Into the Active Sitesupporting

confidence: 62%

Mechanism of transcriptional stalling at cisplatin-damaged DNA

Damsma

Alt

Brueckner

et al. 2007

Nat Struct Mol Biol

160

133

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“…The stability of RNAPII complexes arrested at sites of DNA damage has been monitored by their response to the elongation factor SII (8,33). SII activates a cryptic ribonuclease activity of RNAPII and promotes RNAPII read-through of pause sequences during transcription (34). In response to SII, stalled RNAPII complexes reverse translocate on the template, and RNAPII cleaves off a short sequence from the 3Ј end of the RNA transcript.…”

Section: M1dg and Pdg Adducts Arrest Mammalian Rnapii Translocationmentioning

confidence: 99%

Malondialdehyde adducts in DNA arrest transcription by T7 RNA polymerase and mammalian RNA polymerase II

Cline

Riggins

Tornaletti

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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“…Pol II maintains the bubble, selects nucleoside triphosphates (NTPs) in a template-directed manner, synthesizes RNA, translocates along the DNA and separates RNA from DNA at the upstream end of the hybrid. Pol II also has weak RNA nuclease activity, which is stimulated by the transcript cleavage factor TFIIS [2,3]. Recent reviews of Pol II structure concentrate on TFIIS function and transcription initiation [4][5][6][7].…”

Section: Introductionmentioning

confidence: 99%

The dynamic machinery of mRNA elongation

Armache

Kettenberger

Cramer

2005

Current Opinion in Structural Biology

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Two complementary X-ray studies of the interaction between RNA polymerase II and nucleic acids have improved our understanding of mRNA elongation. These studies suggest how RNA polymerase II unwinds DNA, how it separates the RNA product from the DNA template and how it incorporates nucleoside triphosphate (NTP) substrates into the growing RNA chain. The tunable polymerase active center apparently allows repositioning of a catalytic metal ion, rotation of NTPs before their incorporation, RNA repositioning by a transcript cleavage factor, and modulation of enzyme activity by a bacterial small molecule regulator and its protein cofactor.

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Transcription elongation factor SII

Cited by 187 publications

References 93 publications

Mechanism of transcriptional stalling at cisplatin-damaged DNA

Mechanism of transcriptional stalling at cisplatin-damaged DNA

Malondialdehyde adducts in DNA arrest transcription by T7 RNA polymerase and mammalian RNA polymerase II

The dynamic machinery of mRNA elongation

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