Abstract
Escherichia coli RNA polymerase holoenzyme consists of a core enzyme with subunit composition, ββ;’ α2, capable of elongation of transcription and an additional subunit, sigma (σ)that binds to the core enzyme and allows specific promoter recognition and transcription initiation. Primarily two DNA consensus sequences centered around −10 and −35 are included in the process of promoter recognition and opening. The open complex includes a region where base pairing is disrupted over a region of ∼12 bp. When the open complex is formed, the sigma subunit is released and a processive elongation complex is formed (1–3).
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References
Burgess R.R., Travers A.A., Dunn J.J., and Bautz E.K. (1969) Factor stimulating transcription by RNA polymerase.Nature 4, 43–46.
McClure W.R. (1985) Mechanism and control of transcription initiation in prokaryotes. Annu. Rev. Biochem. 54, 171–204.
Von Hippel P.H., Bear D.G., Morgan W.D., and McSwiggen J.A. (1984) Protein-nucleic acid interactions in transcription:a molecular analysis. Annu. Rev. Biochem. 53, 389–446.
Daube S.S. and vonHippel P.H. (1992) Functional transcription elongation complexes from synthetic RNA-DNA bubble duplexes.Science 258, 1320–1324.
Aiyar S.E., Helmann J.D., and deHaseth P.L. (1994) A mismatch bubble in double-stranded DNA suffices to direct precise transcription initiation by Escherichia coli RNA polymerase. J. Biol. Chem. 6, 13,179–13,184.
Cherny D.Y., Belotserkovskii B.P., Frank-Kamenetskii M.D., et al. (1993) DNA unwinding upon strand-displacement binding of a thymine-substituted polyamide to double-stranded DNA. Proc. Natl. Acad. Sci. USA 90, 1667–1670.
Hanvey J.C., Peffer N.J., Bisi J.E., et al.(1992) Antisense and antigene properties of peptide nucleic acids. Science 258, 1481–1485.
Nielsen P.E., Egholm M., Berg R.H., and Buchardt O. (1991) Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide. Science 254, 1497–1500.
Egholm M., Buchardt O., Nielsen P.E., and Berg R.H. (1992) Peptide nucleic acids (PNA).Oligonucleotide analogues with an achiral peptide backbone J. Amer. Chem. Soc. 114, 9677–9678.
Møllegaard N.E., Buchardt O., Egholm M. and Nielsen P.E. (1994) Peptide nucleic acid.DNA strand displacement loops as artificial transcription promoters. Proc. Natl. Acad. Sci. USA 91, 3892–2895.
Wang G., Xu X., Pace B., et al. (1999) Peptide nucleic acid (PNA) binding-mediated induction of human gamma-globin gene expression. Nucleic Acids Res. 27, 2806–2813.
Egholm M., Christensen L., Dueholm K.L., Buchardt O., Coull J., and Nielsen P.E. (1995) Efficient pH-independent sequence-specific DNA binding by pseudoisocytosine-containing bis-PNA. Nucleic Acids Res. 23, 217–222.
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© 2002 Humana Press Inc.
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MΦllegaard, N.E., Nielsen, P.E. (2002). In Vitro Transcription from Peptide Nucleic Acid/DNA Strand Displacement Loops. In: Nielsen, P.E. (eds) Peptide Nucleic Acids. Methods in Molecular Biology, vol 208. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-290-2:249
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DOI: https://doi.org/10.1385/1-59259-290-2:249
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-976-6
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