Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (438K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (27)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Scholz, B.
Right arrow Articles by Bogner, E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Scholz, B.
Right arrow Articles by Bogner, E.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2003, Vol. 31, No. 5 1426-1433
© 2003 Oxford University Press

Identification of the ATP-binding site in the terminase subunit pUL56 of human cytomegalovirus

Brigitte Scholz, Sabine Rechter, John C. Drach1,2, Leroy B. Townsend2 and Elke Bogner*

Institut für Klinische und Molekulare Virologie, Schlossgarten 4, D-91054 Erlangen, Germany, 1 Department of Biologic and Materials Sciences, School of Dentistry and 2 Interdepartmental Graduate Program in Medicinal Chemistry, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA

*To whom correspondence should be addressed. Tel: +49 9131 8522104; Fax: +49 9131 8526493; Email: eebogner{at}viro.med.uni-erlangen.de

Human cytomegalovirus (HCMV) terminase is composed of subunits pUL56 (130 kDa) and pUL89 (~75 kDa), encoded by the UL56 and UL89 genes. In a recent investigation, we demonstrated that the main ATPase activity is associated with the large terminase subunit pUL56. The protein has two putative ATP-binding sites, which were suggested to be composed of the sequence (amino acids 463–470) for ATP-binding site 1 and YNETFGKQ (amino acids 709–716) for the second site. We now demonstrate using a 1.5 kb fragment encoding the C-terminal half of pUL56 that ATP-binding site 1 is not critical for the function, whereas ATP-binding site 2 is required for the enzymatic activity. Mutation G714A in this protein reduced the ATPase activity to ~65% and the double mutation G714A/K715N showed a reduction up to 75%. However, the substitution of E711A revoked the effect of the substitutions. The functional character of the ATP-binding site was demonstrated by transfer of YNETFGKQLSIACLR (709–723) to glutathione-S-transferase (GST). Interestingly, vanadate, an ATPase inhibitor, has the ability to block the ATPase activity of pUL56 as well as of Apyrase, while the antitumor ATP-mimetic agent geldanamycin, did not affect the ATP-binding of pUL56. Furthermore, in contrast to an inactive control compound, the specific HCMV terminase inhibitor BDCRB showed a partial inhibition of the pUL56-specific ATPase activity. Our results clearly demonstrated that (i) the enzymatic activity of the terminase subunit pUL56 could be inhibited by vanadate, (ii) only the ATP-binding site 2 is critical for the pUL56 function and (iii) glycine G714 is an invariant amino acid.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Antimicrob. Agents Chemother.Home page
J.-S. Hwang, R. Schilf, J. C. Drach, L. B. Townsend, and E. Bogner
Susceptibilities of Human Cytomegalovirus Clinical Isolates and Other Herpesviruses to New Acetylated, Tetrahalogenated Benzimidazole D-Ribonucleosides
Antimicrob. Agents Chemother., December 1, 2009; 53(12): 5095 - 5101.
[Abstract] [Full Text] [PDF]

Home page
 J. Gen. Virol.Home page
A. Holzenburg, A. Dittmer, and E. Bogner
Assembly of monomeric human cytomegalovirus pUL104 into portal structures
J. Gen. Virol., October 1, 2009; 90(10): 2381 - 2385.
[Abstract] [Full Text] [PDF]

Home page
 J. Gen. Virol.Home page
J. B. Wang and M. A. McVoy
Mutagenesis of the murine cytomegalovirus M56 terminase gene
J. Gen. Virol., November 1, 2008; 89(11): 2864 - 2868.
[Abstract] [Full Text] [PDF]

Home page
 J. Lipid Res.Home page
Y.-M. Zhang and C. O. Rock
Thematic Review Series: Glycerolipids. Acyltransferases in bacterial glycerophospholipid synthesis
J. Lipid Res., September 1, 2008; 49(9): 1867 - 1874.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
J.-S. Hwang, O. Kregler, R. Schilf, N. Bannert, J. C. Drach, L. B. Townsend, and E. Bogner
Identification of Acetylated, Tetrahalogenated Benzimidazole D-Ribonucleosides with Enhanced Activity against Human Cytomegalovirus
J. Virol., November 1, 2007; 81(21): 11604 - 11611.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y.-J. Lu, F. Zhang, K. D. Grimes, R. E. Lee, and C. O. Rock
Topology and Active Site of PlsY: THE BACTERIAL ACYLPHOSPHATE:GLYCEROL-3-PHOSPHATE ACYLTRANSFERASE
J. Biol. Chem., April 13, 2007; 282(15): 11339 - 11346.
[Abstract] [Full Text] [PDF]

Home page
 Drug Metab. Dispos.Home page
P. L. Lorenzi, C. P. Landowski, A. Brancale, X. Song, L. B. Townsend, J. C. Drach, and G. L. Amidon
N-METHYLPURINE DNA GLYCOSYLASE AND 8-OXOGUANINE DNA GLYCOSYLASE METABOLIZE THE ANTIVIRAL NUCLEOSIDE 2-BROMO-5,6-DICHLORO-1-({beta}-D-RIBOFURANOSYL)BENZIMIDAZOLE
Drug Metab. Dispos., June 1, 2006; 34(6): 1070 - 1077.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
A. Dittmer, J. C. Drach, L. B. Townsend, A. Fischer, and E. Bogner
Interaction of the Putative Human Cytomegalovirus Portal Protein pUL104 with the Large Terminase Subunit pUL56 and Its Inhibition by Benzimidazole-D-Ribonucleosides
J. Virol., December 1, 2005; 79(23): 14660 - 14667.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
M. A. McVoy and D. E. Nixon
Impact of 2-Bromo-5,6-Dichloro-1-{beta}-D-Ribofuranosyl Benzimidazole Riboside and Inhibitors of DNA, RNA, and Protein Synthesis on Human Cytomegalovirus Genome Maturation
J. Virol., September 1, 2005; 79(17): 11115 - 11127.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
P. L. Lorenzi, C. P. Landowski, X. Song, K. Z. Borysko, J. M. Breitenbach, J. S. Kim, J. M. Hilfinger, L. B. Townsend, J. C. Drach, and G. L. Amidon
Amino Acid Ester Prodrugs of 2-Bromo-5,6-dichloro-1-({beta}-D-ribofuranosyl)benzimidazole Enhance Metabolic Stability in Vitro and in Vivo
J. Pharmacol. Exp. Ther., August 1, 2005; 314(2): 883 - 890.
[Abstract] [Full Text] [PDF]

Home page
 Antimicrob. Agents Chemother.Home page
D. L. Evers, G. Komazin, R. G. Ptak, D. Shin, B. T. Emmer, L. B. Townsend, and J. C. Drach
Inhibition of Human Cytomegalovirus Replication by Benzimidazole Nucleosides Involves Three Distinct Mechanisms
Antimicrob. Agents Chemother., October 1, 2004; 48(10): 3918 - 3927.
[Abstract] [Full Text] [PDF]

Home page
 Antimicrob. Agents Chemother.Home page
M. R. Underwood, R. G. Ferris, D. W. Selleseth, M. G. Davis, J. C. Drach, L. B. Townsend, K. K. Biron, and F. L. Boyd
Mechanism of Action of the Ribopyranoside Benzimidazole GW275175X against Human Cytomegalovirus
Antimicrob. Agents Chemother., May 1, 2004; 48(5): 1647 - 1651.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
D. E. Nixon and M. A. McVoy
Dramatic Effects of 2-Bromo-5,6-Dichloro-1-{beta}-D-Ribofuranosyl Benzimidazole Riboside on the Genome Structure, Packaging, and Egress of Guinea Pig Cytomegalovirus
J. Virol., February 15, 2004; 78(4): 1623 - 1635.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
G. Komazin, L. B. Townsend, and J. C. Drach
Role of a Mutation in Human Cytomegalovirus Gene UL104 in Resistance to Benzimidazole Ribonucleosides
J. Virol., January 15, 2004; 78(2): 710 - 715.
[Abstract] [Full Text] [PDF]

Home page
 Antimicrob. Agents Chemother.Home page
S. L. Williams, C. B. Hartline, N. L. Kushner, E. A. Harden, D. J. Bidanset, J. C. Drach, L. B. Townsend, M. R. Underwood, K. K. Biron, and E. R. Kern
In Vitro Activities of Benzimidazole D- and L-Ribonucleosides against Herpesviruses
Antimicrob. Agents Chemother., July 1, 2003; 47(7): 2186 - 2192.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.