HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Pisani, F. M. Articles by Rossi, M. Search for Related Content PubMed PubMed Citation Articles by Pisani, F. M. Articles by Rossi, M. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 269, Issue 11, 7887-7892, 03, 1994

Evidence that an archaeal alpha-like DNA polymerase has a modular organization of its associated catalytic activities

FM Pisani and M Rossi
Istituto di Biochimica delle Proteine ed Enzimologia, Consiglio Nazionale delle Ricerche, Naples, Italy.

In this study we report on the evidence that an alpha-like DNA polymerase purified from the thermoacidophilic archaeon Sulfolobus solfataricus has a modular organization of its associated catalytic activities (polymerase and 3'-5' exonuclease). This enzyme, a monomer of about 100 kDa whose complete primary structure is available, has a protease hypersensitive site that is likely to be cleaved by the action of endogenous proteases during the purification procedure. As a consequence of that, two proteolytic fragments of about 50 and 40 kDa, in addition to the intact 100-kDa molecular species, can be detected upon SDS-PAGE of highly purified S. solfataricus DNA polymerase samples. The amino-terminal microsequence analysis by Edman degradation has revealed that the 50- and the 40-kDa polypeptides correspond to the carboxyl- and the amino-terminal portion of the protein molecule, respectively. Using the bidimensional activity gel assay procedure, recently described by Longley and Mosbaugh (Longley, M. J., and Mosbaugh, D. W. (1991) Biochemistry 30, 2655-2664), we have demonstrated that the 50-kDa fragment retains a Mg(2+)-dependent DNA polymerizing activity, whereas the 40-kDa polypeptide is able to catalyze the excision of mispaired nucleotides at the 3'-OH terminus of a primer/template DNA substrate in the presence of Mn2+ ions. On the other hand, the 100-kDa protein possess both activities. To date, this is the first report indicating, on the basis of direct functional data, that the polymerization and the 3'-5' exonuclease activity of a family B DNA polymerase can be ascribed to physically distinct modules of the enzyme molecule.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				T. Aucelli, P. Contursi, M. Girfoglio, M. Rossi, and R. Cannio A spreadable, non-integrative and high copy number shuttle vector for Sulfolobus solfataricus based on the genetic element pSSVx from Sulfolobus islandicus Nucleic Acids Res., October 18, 2006; 34(17): e114 - e114. [Abstract] [Full Text] [PDF]

				Y. Asami, M. Murakami, M. Shimizu, F. M. Pisani, I. Hayata, and T. Nohmi Visualization of the interaction between archaeal DNA polymerase and uracil-containing DNA by atomic force microscopy. Genes Cells, January 1, 2006; 11(1): 3 - 11. [Abstract] [Full Text] [PDF]

				K. Komori and Y. Ishino Functional interdependence of DNA polymerizing and 3'->5' exonucleolytic activities in Pyrococcus furiosus DNA polymerase I Protein Eng. Des. Sel., January 1, 2000; 13(1): 41 - 47. [Abstract] [Full Text] [PDF]

				I. K. O. Cann, S. Ishino, N. Nomura, Y. Sako, and Y. Ishino Two Family B DNA Polymerases from Aeropyrum pernix, an Aerobic Hyperthermophilic Crenarchaeote J. Bacteriol., October 1, 1999; 181(19): 5984 - 5992. [Abstract] [Full Text]

				X.-m. Yang and C. C. Richardson Structural and Functional Organization of the DNA Polymerase of Bacteriophage T7 J. Biol. Chem., September 27, 1996; 271(39): 24207 - 24212. [Abstract] [Full Text] [PDF]