1S0V

Structural basis for substrate selection by T7 RNA polymerase

Summary for 1S0V

• Entry DOI: 10.2210/pdb1s0v/pdb
• Related: 1H38
• Descriptor: 5'-D(*G*GP*GP*AP*AP*TP*CP*GP*AP*TP*AP*TP*CP*GP*CP*CP*GP*C)-3', 5'-R(*AP*AP*CP*U*GP*CP*GP*GP*CP*GP*AP*U)-3', 5'-D(*GP*TP*CP*GP*AP*TP*TP*CP*CP*C)-3', ... (7 entities in total)
• Functional Keywords: t7 rna polymerase, dna, rna, riken structural genomics/proteomics initiative, rsgi, structural genomics, transferase-dna-rna complex, transferase-dna-rna hybrid complex, transferase/dna-rna hybrid
• Biological source: Enterobacteria phage T7
• Total number of polymer chains: 16
• Total formula weight: 447707.85

Authors

Temiakov, D.,Patlan, V.,Anikin, M.,McAllister, W.T.,Yokoyama, S.,Vassylyev, D.G.,RIKEN Structural Genomics/Proteomics Initiative (RSGI) (deposition date: 2004-01-05, release date: 2004-02-24, Last modification date: 2023-08-23)

Primary citation

Temiakov, D.,Patlan, V.,Anikin, M.,McAllister, W.T.,Yokoyama, S.,Vassylyev, D.G.
Structural basis for substrate selection by t7 RNA polymerase.
Cell(Cambridge,Mass.), 116:381-391, 2004

PubMed Abstract: The mechanism by which nucleotide polymerases select the correct substrate is of fundamental importance to the fidelity of DNA replication and transcription. During the nucleotide addition cycle, pol I DNA polymerases undergo the transition from a catalytically inactive "open" to an active "closed" conformation. All known determinants of substrate selection are associated with the "closed" state. To elucidate if this mechanism is conserved in homologous single subunit RNA polymerases (RNAPs), we have determined the structure of T7 RNAP elongation complex with the incoming substrate analog. Surprisingly, the substrate specifically binds to RNAP in the "open" conformation, where it is base paired with the acceptor template base, while Tyr639 provides discrimination of ribose versus deoxyribose substrates. The structure therefore suggests a novel mechanism, in which the substrate selection occurs prior to the isomerization to the catalytically active conformation. Modeling of multisubunit RNAPs suggests that this mechanism might be universal for all RNAPs.

PubMed: 15016373
DOI: 10.1016/S0092-8674(04)00059-5

Experimental method

X-RAY DIFFRACTION (3.2 Å)