• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
J Clin Invest. Feb 15, 1996; 97(4): 957–962.
PMCID: PMC507141

Induction of Fanconi anemia cellular phenotype in human 293 cells by overexpression of a mutant FAC allele.

Abstract

The polypeptide encoded by the Fanconi anemia (FA) complementation group C gene, FAC, binds to a group of cytoplasmic proteins in vitro and may form a multimeric complex. A known mutant allele of FAC resulting from the substitution of Pro for Leu at codon 554 fails to correct the sensitivity of FA group C cells to mitomycin C. We reasoned that overexpression of the mutant protein in a wild-type cellular background might induce the FA phenotype by competing with endogenous FAC for binding to the accessory proteins. After stable transfection of 293 cells with wild-type and a mutant FAC allele containing the L554P substitution, four independent clones that expressed four-to-fifteen fold higher levels of transcript from the mutant transgene relative to the endogenous FAC gene showed hypersensitivity to mitomycin C. By contrast, both parental and FAC-overexpressing cells maintained their relative resistance to mitomycin C. No differences in the biosynthesis, subcellular localization and protein interactions of the normal and mutant proteins were detected. The induction of the FA phenotype in this system is compatible with the competition hypothesis and provides support for a functional role of the FAC-binding proteins in vivo.

Full Text

The Full Text of this article is available as a PDF (260K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Liu JM, Buchwald M, Walsh CE, Young NS. Fanconi anemia and novel strategies for therapy. Blood. 1994 Dec 15;84(12):3995–4007. [PubMed]
  • Setlow RB. Repair deficient human disorders and cancer. Nature. 1978 Feb 23;271(5647):713–717. [PubMed]
  • Strathdee CA, Duncan AM, Buchwald M. Evidence for at least four Fanconi anaemia genes including FACC on chromosome 9. Nat Genet. 1992 Jun;1(3):196–198. [PubMed]
  • Joenje H, Lo ten Foe JR, Oostra AB, van Berkel CG, Rooimans MA, Schroeder-Kurth T, Wegner RD, Gille JJ, Buchwald M, Arwert F. Classification of Fanconi anemia patients by complementation analysis: evidence for a fifth genetic subtype. Blood. 1995 Sep 15;86(6):2156–2160. [PubMed]
  • Yamashita T, Barber DL, Zhu Y, Wu N, D'Andrea AD. The Fanconi anemia polypeptide FACC is localized to the cytoplasm. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6712–6716. [PMC free article] [PubMed]
  • Youssoufian H. Localization of Fanconi anemia C protein to the cytoplasm of mammalian cells. Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):7975–7979. [PMC free article] [PubMed]
  • Strathdee CA, Gavish H, Shannon WR, Buchwald M. Cloning of cDNAs for Fanconi's anaemia by functional complementation. Nature. 1992 Apr 30;356(6372):763–767. [PubMed]
  • Murer-Orlando M, Llerena JC, Jr, Birjandi F, Gibson RA, Mathew CG. FACC gene mutations and early prenatal diagnosis of Fanconi's anaemia. Lancet. 1993 Sep 11;342(8872):686–686. [PubMed]
  • Whitney MA, Saito H, Jakobs PM, Gibson RA, Moses RE, Grompe M. A common mutation in the FACC gene causes Fanconi anaemia in Ashkenazi Jews. Nat Genet. 1993 Jun;4(2):202–205. [PubMed]
  • Verlander PC, Lin JD, Udono MU, Zhang Q, Gibson RA, Mathew CG, Auerbach AD. Mutation analysis of the Fanconi anemia gene FACC. Am J Hum Genet. 1994 Apr;54(4):595–601. [PMC free article] [PubMed]
  • Gibson RA, Hajianpour A, Murer-Orlando M, Buchwald M, Mathew CG. A nonsense mutation and exon skipping in the Fanconi anaemia group C gene. Hum Mol Genet. 1993 Jun;2(6):797–799. [PubMed]
  • Gavish H, dos Santos CC, Buchwald M. A Leu554-to-Pro substitution completely abolishes the functional complementing activity of the Fanconi anemia (FACC) protein. Hum Mol Genet. 1993 Feb;2(2):123–126. [PubMed]
  • Youssoufian H, Auerbach AD, Verlander PC, Steimle V, Mach B. Identification of cytosolic proteins that bind to the Fanconi anemia complementation group C polypeptide in vitro. Evidence for a multimeric complex. J Biol Chem. 1995 Apr 28;270(17):9876–9882. [PubMed]
  • Graham FL, Smiley J, Russell WC, Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. [PubMed]
  • Davies MV, Kaufman RJ. The sequence context of the initiation codon in the encephalomyocarditis virus leader modulates efficiency of internal translation initiation. J Virol. 1992 Apr;66(4):1924–1932. [PMC free article] [PubMed]
  • Frangioni JV, Neel BG. Solubilization and purification of enzymatically active glutathione S-transferase (pGEX) fusion proteins. Anal Biochem. 1993 Apr;210(1):179–187. [PubMed]
  • Westerveld A, Hoeijmakers JH, van Duin M, de Wit J, Odijk H, Pastink A, Wood RD, Bootsma D. Molecular cloning of a human DNA repair gene. Nature. 1984 Aug 2;310(5976):425–429. [PubMed]
  • Arlett CF, Lehmann AR. Human disorders showing increased sensitivity to the induction of genetic damage. Annu Rev Genet. 1978;12:95–115. [PubMed]
  • Belt PB, van Oosterwijk MF, Odijk H, Hoeijmakers JH, Backendorf C. Induction of a mutant phenotype in human repair proficient cells after overexpression of a mutated human DNA repair gene. Nucleic Acids Res. 1991 Oct 25;19(20):5633–5637. [PMC free article] [PubMed]
  • van Vuuren AJ, Appeldoorn E, Odijk H, Yasui A, Jaspers NG, Bootsma D, Hoeijmakers JH. Evidence for a repair enzyme complex involving ERCC1 and complementing activities of ERCC4, ERCC11 and xeroderma pigmentosum group F. EMBO J. 1993 Sep;12(9):3693–3701. [PMC free article] [PubMed]
  • Li L, Elledge SJ, Peterson CA, Bales ES, Legerski RJ. Specific association between the human DNA repair proteins XPA and ERCC1. Proc Natl Acad Sci U S A. 1994 May 24;91(11):5012–5016. [PMC free article] [PubMed]
  • Li L, Peterson CA, Lu X, Legerski RJ. Mutations in XPA that prevent association with ERCC1 are defective in nucleotide excision repair. Mol Cell Biol. 1995 Apr;15(4):1993–1998. [PMC free article] [PubMed]
  • van Vuuren AJ, Appeldoorn E, Odijk H, Humbert S, Moncollin V, Eker AP, Jaspers NG, Egly JM, Hoeijmakers JH. Partial characterization of the DNA repair protein complex, containing the ERCC1, ERCC4, ERCC11 and XPF correcting activities. Mutat Res. 1995 Jul;337(1):25–39. [PubMed]
  • Gal A, Artlich A, Ludwig M, Niemeyer G, Olek K, Schwinger E, Schinzel A. Pro-347-Arg mutation of the rhodopsin gene in autosomal dominant retinitis pigmentosa. Genomics. 1991 Oct;11(2):468–470. [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links