2.50
Hdl Handle:
http://hdl.handle.net/10033/592867
Title:
The 3D structure of Kaposi sarcoma herpesvirus LANA C-terminal domain bound to DNA.
Authors:
Hellert, Jan; Weidner-Glunde, Magdalena; Krausze, Joern; Lünsdorf, Heinrich; Ritter, Christiane; Schulz, Thomas F; Lührs, Thorsten
Abstract:
Kaposi sarcoma herpesvirus (KSHV) persists as a latent nuclear episome in dividing host cells. This episome is tethered to host chromatin to ensure proper segregation during mitosis. For duplication of the latent genome, the cellular replication machinery is recruited. Both of these functions rely on the constitutively expressed latency-associated nuclear antigen (LANA) of the virus. Here, we report the crystal structure of the KSHV LANA DNA-binding domain (DBD) in complex with its high-affinity viral target DNA, LANA binding site 1 (LBS1), at 2.9 Å resolution. In contrast to homologous proteins such as Epstein-Barr virus nuclear antigen 1 (EBNA-1) of the related γ-herpesvirus Epstein-Barr virus, specific DNA recognition by LANA is highly asymmetric. In addition to solving the crystal structure, we found that apart from the two known LANA binding sites, LBS1 and LBS2, LANA also binds to a novel site, denoted LBS3. All three sites are located in a region of the KSHV terminal repeat subunit previously recognized as a minimal replicator. Moreover, we show that the LANA DBD can coat DNA of arbitrary sequence by virtue of a characteristic lysine patch, which is absent in EBNA-1 of the Epstein-Barr virus. Likely, these higher-order assemblies involve the self-association of LANA into supermolecular spirals. One such spiral assembly was solved as a crystal structure of 3.7 Å resolution in the absence of DNA. On the basis of our data, we propose a model for the controlled nucleation of higher-order LANA oligomers that might contribute to the characteristic subnuclear KSHV microdomains ("LANA speckles"), a hallmark of KSHV latency.
Citation:
The 3D structure of Kaposi sarcoma herpesvirus LANA C-terminal domain bound to DNA. 2015, 112 (21):6694-9 Proc. Natl. Acad. Sci. U.S.A.
Journal:
Proceedings of the National Academy of Sciences of the United States of America
Issue Date:
26-May-2015
URI:
http://hdl.handle.net/10033/592867
DOI:
10.1073/pnas.1421804112
PubMed ID:
25947153
Type:
Article
Language:
en
ISSN:
1091-6490
Appears in Collections:
publications of the department infection genetics (INFG)

Full metadata record

DC FieldValue Language
dc.contributor.authorHellert, Janen
dc.contributor.authorWeidner-Glunde, Magdalenaen
dc.contributor.authorKrausze, Joernen
dc.contributor.authorLünsdorf, Heinrichen
dc.contributor.authorRitter, Christianeen
dc.contributor.authorSchulz, Thomas Fen
dc.contributor.authorLührs, Thorstenen
dc.date.accessioned2016-01-05T16:42:19Zen
dc.date.available2016-01-05T16:42:19Zen
dc.date.issued2015-05-26en
dc.identifier.citationThe 3D structure of Kaposi sarcoma herpesvirus LANA C-terminal domain bound to DNA. 2015, 112 (21):6694-9 Proc. Natl. Acad. Sci. U.S.A.en
dc.identifier.issn1091-6490en
dc.identifier.pmid25947153en
dc.identifier.doi10.1073/pnas.1421804112en
dc.identifier.urihttp://hdl.handle.net/10033/592867en
dc.description.abstractKaposi sarcoma herpesvirus (KSHV) persists as a latent nuclear episome in dividing host cells. This episome is tethered to host chromatin to ensure proper segregation during mitosis. For duplication of the latent genome, the cellular replication machinery is recruited. Both of these functions rely on the constitutively expressed latency-associated nuclear antigen (LANA) of the virus. Here, we report the crystal structure of the KSHV LANA DNA-binding domain (DBD) in complex with its high-affinity viral target DNA, LANA binding site 1 (LBS1), at 2.9 Å resolution. In contrast to homologous proteins such as Epstein-Barr virus nuclear antigen 1 (EBNA-1) of the related γ-herpesvirus Epstein-Barr virus, specific DNA recognition by LANA is highly asymmetric. In addition to solving the crystal structure, we found that apart from the two known LANA binding sites, LBS1 and LBS2, LANA also binds to a novel site, denoted LBS3. All three sites are located in a region of the KSHV terminal repeat subunit previously recognized as a minimal replicator. Moreover, we show that the LANA DBD can coat DNA of arbitrary sequence by virtue of a characteristic lysine patch, which is absent in EBNA-1 of the Epstein-Barr virus. Likely, these higher-order assemblies involve the self-association of LANA into supermolecular spirals. One such spiral assembly was solved as a crystal structure of 3.7 Å resolution in the absence of DNA. On the basis of our data, we propose a model for the controlled nucleation of higher-order LANA oligomers that might contribute to the characteristic subnuclear KSHV microdomains ("LANA speckles"), a hallmark of KSHV latency.en
dc.language.isoenen
dc.subject.meshAmino Acid Sequenceen
dc.subject.meshAntigens, Viralen
dc.subject.meshBase Sequenceen
dc.subject.meshBinding Sitesen
dc.subject.meshCrystallography, X-Rayen
dc.subject.meshDNA, Viralen
dc.subject.meshDNA-Binding Proteinsen
dc.subject.meshHerpesvirus 8, Humanen
dc.subject.meshHumansen
dc.subject.meshModels, Molecularen
dc.subject.meshMolecular Sequence Dataen
dc.subject.meshMutagenesis, Site-Directeden
dc.subject.meshNuclear Proteinsen
dc.subject.meshProtein Structure, Quaternaryen
dc.subject.meshProtein Structure, Tertiaryen
dc.subject.meshScattering, Small Angleen
dc.subject.meshStatic Electricityen
dc.subject.meshX-Ray Diffractionen
dc.titleThe 3D structure of Kaposi sarcoma herpesvirus LANA C-terminal domain bound to DNA.en
dc.typeArticleen
dc.identifier.journalProceedings of the National Academy of Sciences of the United States of Americaen
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