Structural analysis of human Orc6 protein reveals a homology with transcription factor TFIIB

The Origin Recognition Complex (ORC) is a six-subunit protein important for the initiation of DNA replication in eukaryotic cells. Orc6 is the smallest and the least conserved among ORC subunits. It is required for the DNA replication but also has a function in cytokinesis in metazoan species, howev...

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Vydáno v:Proceedings of the National Academy of Sciences - PNAS Ročník 108; číslo 18; s. 7373
Hlavní autoři: Liu, Shixuan, Balasov, Maxim, Wang, Hongfei, Wu, Lijie, Chesnokov, Igor N, Liu, Yingfang
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States 03.05.2011
Témata:
Amino Acid Sequence
Animals
Bromodeoxyuridine
Chromatography, Gel
Cloning, Molecular
Crystallization
DNA Replication - genetics
Drosophila
Electrophoresis, Polyacrylamide Gel
Green Fluorescent Proteins
Humans
Models, Molecular
Molecular Sequence Data
Origin Recognition Complex - chemistry
Origin Recognition Complex - genetics
Protein Binding
Protein Conformation
Sequence Alignment
Sequence Homology
Transcription Factor TFIIB - genetics
Xenopus
ISSN:1091-6490, 1091-6490
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Shrnutí:The Origin Recognition Complex (ORC) is a six-subunit protein important for the initiation of DNA replication in eukaryotic cells. Orc6 is the smallest and the least conserved among ORC subunits. It is required for the DNA replication but also has a function in cytokinesis in metazoan species, however, the mechanisms of Orc6 action in these processes are not clear. Here we report a structure of the middle domain of human Orc6. This domain has an overall fold similar to the corresponding helical domain of transcription factor TFIIB. Based on these findings, a model of Orc6 binding to DNA is produced. We have identified amino acids of Orc6 which are directly involved in DNA binding. Alterations of these amino acids abolish DNA binding ability of Orc6 and also result in reduced levels of DNA replication in vitro and in cultured cells. Our data indicate that Orc6 is one of the DNA binding subunits of ORC in metazoan species. We propose that Orc6 may participate in positioning of ORC at the origins of DNA replication similar to the role of TFIIB in positioning transcription preinitiation complex at the promoter.
Bibliografie:ObjectType-Article-2
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ISSN:1091-6490
1091-6490
DOI:10.1073/pnas.1013676108