Defective FANCI binding by a fanconi anemia-related FANCD2 mutant

Koichi Sato; Masamichi Ishiai; Minoru Takata; Hitoshi Kurumizaka

doi:10.1371/journal.pone.0114752

Defective FANCI binding by a fanconi anemia-related FANCD2 mutant

PLoS One. 2014 Dec 9;9(12):e114752. doi: 10.1371/journal.pone.0114752. eCollection 2014.

Authors

Koichi Sato¹, Masamichi Ishiai², Minoru Takata², Hitoshi Kurumizaka¹

Affiliations

¹ Laboratory of Structural Biology, Graduate School of Advanced Science & Engineering, Waseda University, Tokyo, Japan.
² Laboratory of DNA Damage Signaling, Department of Late Effects Studies, Radiation Biology Center, Kyoto University, Kyoto, Japan.

Abstract

FANCD2 is a product of one of the genes associated with Fanconi anemia (FA), a rare recessive disease characterized by bone marrow failure, skeletal malformations, developmental defects, and cancer predisposition. FANCD2 forms a complex with FANCI (ID complex) and is monoubiquitinated, which facilitates the downstream interstrand crosslink (ICL) repair steps, such as ICL unhooking and nucleolytic end resection. In the present study, we focused on the chicken FANCD2 (cFANCD2) mutant harboring the Leu234 to Arg (L234R) substitution. cFANCD2 L234R corresponds to the human FANCD2 L231R mutation identified in an FA patient. We found that cFANCD2 L234R did not complement the defective ICL repair in FANCD2-/- DT40 cells. Purified cFANCD2 L234R did not bind to chicken FANCI, and its monoubiquitination was significantly deficient, probably due to the abnormal ID complex formation. In addition, the histone chaperone activity of cFANCD2 L234R was also defective. These findings may explain some aspects of Fanconi anemia pathogenesis by a FANCD2 missense mutation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Chickens
DNA Damage*
DNA Repair
Fanconi Anemia / genetics*
Fanconi Anemia / metabolism
Fanconi Anemia Complementation Group D2 Protein / genetics*
Humans
Molecular Sequence Data
Mutation, Missense / genetics*
Protein Binding
Sequence Homology, Amino Acid
Ubiquitin-Conjugating Enzymes / metabolism
Ubiquitination

Substances

Fanconi Anemia Complementation Group D2 Protein
UBE2T protein, human
Ubiquitin-Conjugating Enzymes

Supplementary concepts

Fanconi Anemia, Complementation Group I

Grants and funding

This work was supported in part by JSPS KAKENHI Grant Numbers 25250023 [to H.K.], 24310042 [to M.T.], 26281021 [to M.I.], and 26830128 [to K.S.], and by MEXT KAKENHI Grant Numbers 25116002 [to H.K.], 23114010 [to M.T.], and 25131706 [to M.I]. H.K. was also supported by the Waseda Research Institute for Science and Engineering and Waseda University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.