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FANCA has intrinsic affinity to nucleic acids with preference for single‐stranded forms
Author(s) -
Yuan Fenghua,
Qian Liangyue,
Zhao Xinliang,
Liu Jesse Y,
Song Limin,
D'Urso Gennaro,
Jain Chaitanya,
Zhang Yanbin
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.539.15
Subject(s) - fanca , nucleic acid , dna , fanconi anemia , rna , biology , microbiology and biotechnology , biochemistry , gene , dna repair
Fanconi anemia complementation group A (FANCA) gene is one of the 15 disease‐causing genes and has been found to be mutated in ~60% of Fanconi anemia patients. Using purified protein, we report that human FANCA has intrinsic affinity to nucleic acids. FANCA binds to both single‐stranded and double‐stranded DNA, however, its affinity to single stranded DNA is significantly higher than to double‐stranded DNA in an electrophoretic mobility shift assay. FANCA also binds to RNA with an intriguingly higher affinity than its DNA counterpart. FANCA requires a certain length of nucleic acids for optimal binding. Using DNA and RNA ladders, we determined that the minimum numbers of nucleotides required for FANCA recognition are ~30 for both DNA and RNA. By testing the affinity between FANCA and a variety of DNA structures, we found that a 5′ flap or tail on DNA facilitates its interaction with FANCA. A patient‐derived FANCA truncation mutant, Q772X, has diminished affinity to both DNA and RNA. On the contrary, the complementing C‐terminal fragment of Q772X, C772‐1455, retains the differentiated nucleic acids binding activity (RNA>ssDNA>dsDNA), indicating that the nucleic acids binding domain of FANCA is primarily located at its C‐terminus where most disease causing mutations are found.