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

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Lin, H. Articles by Black, L. W. Search for Related Content PubMed PubMed Citation Articles by Lin, H. Articles by Black, L. W. Social Bookmarking                      What's this?

Volume 272, Number 6, Issue of February 7, 1997 pp. 3495-3501
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Purification and Characterization of the Small Subunit of Phage T4 Terminase, gp16, Required for DNA Packaging

(Received for publication, August 7, 1996, and in revised form, November 13, 1996)

Hsingchi Lin , Martha N. Simon ^¶ and Lindsay W. Black

From the  Department of Biochemistry and the Molecular and Cell Biology Graduate Program, University of Maryland Medical School, Baltimore, Maryland 21201-1503 and the ^¶ Biology Department, Brookhaven National Laboratory, Upton, New York 11973

Phage T4 terminase is an enzyme that binds to the portal protein of proheads and cuts and packages concatemeric DNA. The T4 terminase is composed of two subunits, gene products (gp) 16 and 17. The role of the small subunit, gp16, in T4 DNA packaging is not well characterized. We developed a new purification procedure to obtain large quantities of purified gp16 from an overexpression vector. The pure protein is found in two molecular weight forms, due to specific C-terminal truncation, displays in vitro packaging activity, and binds but does not hydrolyze ATP. gp16 forms specific oligomers, rings, and side-by-side double rings, as judged by native polyacrylamide gel electrophoresis and scanning transmission electron microscopy measurements. The single ring contains about eight monomers, and the rings have a diameter of about 8 nm with a central hole of about 2 nm. A DNA-binding helix-turn-helix motif close to the N terminus of gp16 is predicted. The oligomers do not bind to DNA, but following denaturation and renaturation in the presence of DNA, binding can be demonstrated by gel shift and filter binding assays. gp16 binds to double-stranded DNA but not single-stranded DNA, and appears to bind preferentially to a gene 16-containing DNA sequence.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				L. W. Black and G. Peng Mechanistic Coupling of Bacteriophage T4 DNA Packaging to Components of the Replication-dependent Late Transcription Machinery J. Biol. Chem., September 1, 2006; 281(35): 25635 - 25643. [Abstract] [Full Text] [PDF]

				M. S. Mitchell and V. B. Rao Functional Analysis of the Bacteriophage T4 DNA-packaging ATPase Motor J. Biol. Chem., January 6, 2006; 281(1): 518 - 527. [Abstract] [Full Text] [PDF]

				F. Xiao, W.-D. Moll, S. Guo, and P. Guo Binding of pRNA to the N-terminal 14 amino acids of connector protein of bacteriophage phi29 Nucleic Acids Res., May 10, 2005; 33(8): 2640 - 2649. [Abstract] [Full Text] [PDF]

				S. Kanamaru, K. Kondabagil, M. G. Rossmann, and V. B. Rao The Functional Domains of Bacteriophage T4 Terminase J. Biol. Chem., September 24, 2004; 279(39): 40795 - 40801. [Abstract] [Full Text] [PDF]

				E. S. Miller, E. Kutter, G. Mosig, F. Arisaka, T. Kunisawa, and W. Ruger Bacteriophage T4 Genome Microbiol. Mol. Biol. Rev., March 1, 2003; 67(1): 86 - 156. [Abstract] [Full Text] [PDF]

				R. G. Baumann and L. W. Black Isolation and Characterization of T4 Bacteriophage gp17 Terminase, a Large Subunit Multimer with Enhanced ATPase Activity J. Biol. Chem., February 7, 2003; 278(7): 4618 - 4627. [Abstract] [Full Text] [PDF]

				M. S. Mitchell, S. Matsuzaki, S. Imai, and V. B. Rao Sequence analysis of bacteriophage T4 DNA packaging/terminase genes 16 and 17 reveals a common ATPase center in the large subunit of viral terminases Nucleic Acids Res., September 15, 2002; 30(18): 4009 - 4021. [Abstract] [Full Text] [PDF]

				H. Scheffczik, C. G. W. Savva, A. Holzenburg, L. Kolesnikova, and E. Bogner The terminase subunits pUL56 and pUL89 of human cytomegalovirus are DNA-metabolizing proteins with toroidal structure Nucleic Acids Res., April 1, 2002; 30(7): 1695 - 1703. [Abstract] [Full Text] [PDF]

				G. Leffers and V. B. Rao Biochemical Characterization of an ATPase Activity Associated with the Large Packaging Subunit gp17 from Bacteriophage T4 J. Biol. Chem., November 17, 2000; 275(47): 37127 - 37136. [Abstract] [Full Text] [PDF]

				A. Gual, A. G. Camacho, and J. C. Alonso Functional Analysis of the Terminase Large Subunit, G2P, of Bacillus subtilis Bacteriophage SPP1 J. Biol. Chem., November 3, 2000; 275(45): 35311 - 35319. [Abstract] [Full Text] [PDF]