Document Detail


Molecular flexibility and discontinuous translocation of a non-templated polymerase.
MedLine Citation:
PMID:  15033355     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Little is known regarding the translocation of non-templated nucleic acid polymerases with respect to single-stranded primers. VP55, the vaccinia virus poly(A) polymerase, translocates as it processively adds a approximately 3-7 adenylate tail to primers possessing only three ribouridylate residues (as an (rU)(2)-N(15)-rU motif), and a approximately 25-30 adenylate tail to primers that are more U-rich. Here, three models were addressed for the translocation of VP55 with respect to its primer, namely: (a) rigid protein/rigid nucleic acid; (b) flexible protein/rigid nucleic acid; (c) rigid protein/flexible nucleic acid. Analysis of free and covalently VP55-attached primers favored either (b) or a version of (c) incorporating a passive steric block, and suggested two regions of relative motion between polymerase and primer. Inclusion of a 6nt uridylate-rich patch at the primer 3' end switched the polymerase from approximately 3-7 nt to approximately 25-30 nt tail addition without affecting initial binding affinity. By synthesizing this patch as a (rU/dC) pool, discontinuous polymerase movements could be detected.
Authors:
L Johnson; S Liu; P D Gershon
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of molecular biology     Volume:  337     ISSN:  0022-2836     ISO Abbreviation:  J. Mol. Biol.     Publication Date:  2004 Apr 
Date Detail:
Created Date:  2004-03-22     Completed Date:  2004-04-28     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985088R     Medline TA:  J Mol Biol     Country:  England    
Other Details:
Languages:  eng     Pagination:  843-56     Citation Subset:  IM    
Affiliation:
Department of Medical Biochemistry and Genetics, Institute of Biosciences and Technology, Texas A and M University System Health Science Center, Houston, TX 77030, USA.
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MeSH Terms
Descriptor/Qualifier:
Polynucleotide Adenylyltransferase / metabolism*
Protein Binding
Protein Transport
RNA / biosynthesis*
Vaccinia virus / enzymology*,  genetics
Grant Support
ID/Acronym/Agency:
GM51953/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
63231-63-0/RNA; EC 2.7.7.19/Polynucleotide Adenylyltransferase

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