FANCI protein binds to DNA and interacts with FANCD2 to recognize branched structures

Fenghua Yuan, Jimmy El Hokayem, Wen Zhou, Yanbin Zhang

Research output: Contribution to journalArticle

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Abstract

In this study, we report that the purified wild-type FANCI (Fanconi anemia complementation group I) protein directly binds to a variety of DNA substrates. The DNA binding domain roughly encompasses residues 200-1000, as suggested by the truncation study. When co-expressed in insect cells, a small fraction of FANCI forms a stable complex with FANCD2 (Fanconi anemia complementation group D2). Intriguingly, the purified FANCI-FANCD2 complex preferentially binds to the branched DNA structures when compared with either FANCI or FANCD2 alone. Co-immunoprecipitation with purified proteins indicates that FANCI interacts with FANCD2 through its C-terminal amino acid 1001-1328 fragment. Although the C terminus of FANCI is dispensable for direct DNA binding, it seems to be involved in the regulation of DNA binding activity. This notion is further enhanced by two C-terminal point mutations, R1285Q and D1301A, which showed differentiated DNA binding activity. We also demonstrate that FANCI forms discrete nuclear foci in HeLa cells in the absence or presence of exogenous DNA damage. The FANCI foci are colocalized perfectly with FANCD2 and partially with proliferating cell nuclear antigen irrespective of mitomycin C treatment. An increased number of FANCI foci form and become resistant to Triton X extraction in response to mitomycin C treatment. Our data suggest that the FANCI-FANCD2 complex may participate in repair of damaged replication forks through its preferential recognition of branched structures.

Original languageEnglish
Pages (from-to)24443-24452
Number of pages10
JournalJournal of Biological Chemistry
Volume284
Issue number36
DOIs
StatePublished - Sep 4 2009

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Fanconi Anemia Complementation Group Proteins
Fanconi Anemia
DNA
Proteins
Mitomycin
Proliferating Cell Nuclear Antigen
Complementation Group I Fanconi Anemia
HeLa Cells
Immunoprecipitation
Point Mutation
Repair
DNA Damage
Insects
Amino Acids

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

FANCI protein binds to DNA and interacts with FANCD2 to recognize branched structures. / Yuan, Fenghua; El Hokayem, Jimmy; Zhou, Wen; Zhang, Yanbin.

In: Journal of Biological Chemistry, Vol. 284, No. 36, 04.09.2009, p. 24443-24452.

Research output: Contribution to journalArticle

Yuan, Fenghua ; El Hokayem, Jimmy ; Zhou, Wen ; Zhang, Yanbin. / FANCI protein binds to DNA and interacts with FANCD2 to recognize branched structures. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 36. pp. 24443-24452.
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