Structure and inhibition of herpesvirus DNA packaging terminase nuclease domain.
During viral replication, herpesviruses package their DNA into the procapsid by means of the terminase protein complex. In human cytomegalovirus (herpesvirus 5), the terminase is composed of subunits UL89 and UL56. UL89 cleaves the long DNA concatemers into unit-length genomes of appropriate length for encapsidation. We used ESPRIT, a high-throughput screening method, to identify a soluble purifiable fragment of UL89 from a library of 18,432 randomly truncated ul89 DNA constructs. The purified protein was crystallized and its three-dimensional structure was solved. This protein corresponds to the key nuclease domain of the terminase and shows an RNase H/integrase-like fold. We demonstrate that UL89-C has the capacity to process the DNA and that this function is dependent on Mn(2+) ions, two of which are located at the active site pocket. We also show that the nuclease function can be inactivated by raltegravir, a recently approved anti-AIDS drug that targets the HIV integrase.
Nadal
Marta
M
Institute for Research in Biomedicine, Barcelona Science Park, Baldiri Reixac 10, 08028 Barcelona, Spain.
Mas
Philippe J
PJ
Mas
Phillipe J
PJ
Blanco
Alexandre G
AG
Arnan
Carme
C
Solà
Maria
M
Hart
Darren J
DJ
Coll
Miquel
M
D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D003587:Cytomegalovirus / Q000201:enzymology* / Q000235:genetics; D042003:DNA Packaging; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D009154:Mutation; D017434:Protein Structure, Tertiary; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics
D004279:DNA, Viral; C436448:UL89 protein, Cytomegalovirus; D014764:Viral Proteins
Science. 2004 Sep 3;305(5689):1434-7
15284453
Proc Natl Acad Sci U S A. 1976 Aug;73(8):2619-23
183204
Cell. 1995 Jul 28;82(2):209-20
7628012
Science. 2000 Jul 7;289(5476):77-85
10884228
Nat Struct Mol Biol. 2007 Mar;14(3):229-33
17310249
J Virol. 2007 Nov;81(21):11604-11
17728228
J Mol Biol. 2007 Nov 2;373(4):851-65
17870092
Nat Struct Mol Biol. 2008 Nov;15(11):1228-31
18953336
Science. 1994 Dec 23;266(5193):1981-6
7801124
J Virol. 2003 Sep;77(17):9613-21
12915573
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2256-68
15572779
J Mol Biol. 1998 Sep 18;282(2):359-68
9735293
EMBO J. 2006 May 3;25(9):1924-33
16601679
J Virol. 2001 Oct;75(19):9077-86
11533171
Proteins. 2010 May 1;78(6):1520-30
20099308
Cell. 1994 Sep 23;78(6):1063-72
7923356
Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
15299926
Proteins. 1991;11(4):281-96
1758883
J Virol. 1998 Mar;72(3):2259-64
9499084
Biochemistry. 2006 Jul 25;45(29):8855-63
16846228
J Med Chem. 2008 Sep 25;51(18):5843-55
18763751
Antivir Ther. 2007;12(2):217-32
17503664
J Struct Biol. 2010 Oct;172(1):66-74
20206698
J Virol. 2000 Feb;74(3):1587-92
10627574
Cell. 2008 Dec 26;135(7):1251-62
19109896
Nucleic Acids Res. 2002 Apr 1;30(7):1695-703
11917032
J Mol Biol. 2007 Nov 30;374(3):817-36
17945256
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
15572765
Mol Cell. 2007 Mar 23;25(6):943-9
17386269
Nature. 2010 Mar 11;464(7286):232-6
20118915
J Virol. 1998 Jan;72(1):717-25
9420278
J Biol Chem. 2003 Jun 27;278(26):23251-9
12697751
Proc Natl Acad Sci U S A. 1998 Apr 28;95(9):4831-6
9560188
Curr Opin Struct Biol. 2010 Feb;20(1):104-13
20061134
Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):1971-6
20133842
Methods Mol Biol. 2007;364:215-30
17172768
Rev Med Virol. 2002 Mar-Apr;12(2):115-27
11921307
Cell. 2005 Jul 1;121(7):1005-16
15989951
EMBO Rep. 2009 Jun;10(6):592-8
19444313
Mol Cell. 2007 Oct 26;28(2):264-76
17964265
J Virol. 2005 Dec;79(23):14660-7
16282466
Structure. 1995 Feb 15;3(2):131-4
7735828
Nucleic Acids Res. 2003 Jul 1;31(13):3367-9
12824329
Antimicrob Agents Chemother. 2004 Jul;48(7):2760-5
15215146