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ID: 226756.0, MPI für biophysikalische Chemie / Molekulare Biologie (Dr. Thomas M. Jovin)
Scanning force microscopy of the complexes of p53 core domain with supercoiled DNA
Authors:Jett, S. D.; Cherny, D. I.; Subramaniam, V.; Jovin, T. M.
Language:English
Date of Publication (YYYY-MM-DD):2000
Title of Journal:Journal of Molecular Biology
Volume:299
Start Page:585
End Page:592
Copyright:Jahrbuch 2001, Copyright MPG 2001
Review Status:Peer-review
Audience:Not Specified
Intended Educational Use:No
Abstract / Description:We used scanning force microscopy to analyse the interaction of the core domain of the tumor suppressor protein p53 (p53CD, amino acid residues 94 to 312), with supercoiled DNA (scDNA) molecules. The complexes were attached to a mica substrate by the divalent cation spreading technique. p53CD bound to supercoiled plasmid pPGM1 bearing the consensus sequence 5'-AGACATGCCTAGACATGCCT-3' (p53CON) was imaged as a globular complex. Only one such complex was observed with each scDNA molecule. In contrast, binding to supercoiled pBluescript II SK(-) DNA (lacking the consensus sequence) resulted in the appearance of multiple, variable size complexes of various sizes on single DNA molecules. Addition of p53CD to scDNA containing a cruciform-forming (AT)(34) insert resulted in the binding of the protein exclusively at the cruciform. The data presented here suggest that p53CD can form stable specific and non-specific complexes with supercoiled DNA molecules, albeit of variable multimeric organization.
Free Keywords:DNA-protein complexes; p53; supercoiled DNA; cruciform; scanning force microscopy
Last Change of the Resource (YYYY-MM-DD):2005-08-17
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Thomas Jovin / 060
External Affiliations:Institute of Molecular Genetics, Russian Academy of Sciences, Kurchatov's Square, Moscow, Russia
Identifiers:LOCALID:23126
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