Spatial organization of transcription elongation complex in Escherichia coil

Science

Volumn 281, Issue 5375, 1998, Pages 424-428

  Nudler, Evgeny ^a   Gusarov, Ivan ^a   Avetissova, Ekaterina ^b   Kozlov, Maxim ^b   Goldfarb, Alex ^b  

^a NEW YORK UNIVERSITY MEDICAL CENTER   (United States)

^b PUBLIC HEALTH RESEARCH INSTITUTE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

DOUBLE STRANDED DNA; RNA POLYMERASE;

AMINO ACID SEQUENCE; ARTICLE; BACTERIAL GENETICS; BINDING SITE; CROSS LINKING; DNA BINDING; ESCHERICHIA COLI; PRIORITY JOURNAL; PROTEIN DOMAIN; RNA BINDING; RNA SYNTHESIS;

BINDING SITES; DNA, BACTERIAL; DNA-DIRECTED RNA POLYMERASES; ESCHERICHIA COLI; IDOXURIDINE; MODELS, GENETIC; NUCLEIC ACID CONFORMATION; NUCLEIC ACID HETERODUPLEXES; PROTEIN BINDING; RNA, BACTERIAL; TEMPLATES, GENETIC; TRANSCRIPTION, GENETIC; ULTRAVIOLET RAYS;

EID: 0032541021     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.281.5375.424     Document Type: Article

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20. "Azido"-UTP is 5-(p-azidotetrafluoro)benzamido-allil-uridine triphosphate. The complete synthesis of "azido"-UTP will be published elsewhere. 4-thio-UTP was synthesized from 4-thio-UMP (Sigma) as described (14). 5-lodo-2′-deoxyuridine containing DNA oligonucleotide was custom synthesized (Oligos Etc., Wilsonville, OR).
21. 32P]guanosine monophosphate, the cross-linking with 4-thio-UMP was induced by UV irradiation at 365 nm as in (14), the cross-linking with "azido"-UMP was induced by UV irradiation at 308 nm for 5 min, and the cross-linking with 5-iodo-2′-deoxyuridine was performed as in (2). The template used was the T7A1 promoter fragment (153-pb), obtained by polymerase chain reaction with the following transcribed sequence: ATCGAGAGGGACACGGCGAATAGCCATCCCAATCG-AACAGGCCTGCTGGTAATCGCAGGCCTGGAGACTT- GGATCCCCGGGTA.
22. We are grateful to O. Bereschenko for help with the figures, A. Mustaev for advice on cross-linking techniques, and Sergei Borukhov for CreB. This work was supported by NIH grant GM49242 (A.G.) and departmental funds (E.N). E.A. and M.K. are in the graduate programs of the Institute of Molecular Genetics, Russian Academy of Sciences, and the Limnological Institute (Irkutsk, Russia), respectively.