A novel BTB/POZ transcriptional repressor protein interacts with the Fanconi anemia group C protein and PLZF.

Fanconi anemia (FA) is an autosomal recessive cancer susceptibility syndrome. The phenotype includes developmental defects, bone marrow failure, and cell cycle abnormalities. At least eight complementation groups (A-H) exist, and although three of the corresponding complementation group genes have been cloned, they lack recognizable motifs, and their functions are unknown. We have isolated a binding partner for the Fanconi anemia group C protein (FANCC) by yeast two-hybrid screening. We show that the novel gene, FAZF, encodes a 486 amino acid protein containing a conserved amino terminal BTB/POZ protein interaction domain and three C-terminal Krüppel-like zinc fingers. FAZF is homologous to the promyelocytic leukemia zinc finger (PLZF) protein, which has been shown to act as a transcriptional repressor by recruitment of nuclear corepressors (N-CoR, Sin3, and HDAC1 complex). Consistent with a role in FA, BTB/POZ-containing proteins have been implicated in oncogenesis, limb morphogenesis, hematopoiesis, and proliferation. We show that FAZF is a transcriptional repressor that is able to bind to the same DNA target sequences as PLZF. Our data suggest that the FAZF/FANCC interaction maps to a region of FANCC deleted in FA patients with a severe disease phenotype. We also show that FAZF and wild-type FANCC can colocalize in nuclear foci, whereas a patient-derived mutant FANCC that is compromised for nuclear localization cannot. These results suggest that the function of FANCC may be linked to a transcriptional repression pathway involved in chromatin remodeling.

[1]  J. Licht,et al.  Sequence-specific DNA Binding and Transcriptional Regulation by the Promyelocytic Leukemia Zinc Finger Protein* , 1997, The Journal of Biological Chemistry.

[2]  D. Näf,et al.  The Fanconi anaemia proteins, FAA and FAC interact to form a nuclear complex , 1997, Nature Genetics.

[3]  K. Borden,et al.  Two RING Finger Proteins, the Oncoprotein PML and the Arenavirus Z Protein, Colocalize with the Nuclear Fraction of the Ribosomal P Proteins , 1998, Journal of Virology.

[4]  A. D’Andrea,et al.  Nuclear localization of the Fanconi anemia protein FANCC is required for functional activity. , 1999, Blood.

[5]  H. Youssoufian.  Localization of Fanconi anemia C protein to the cytoplasm of mammalian cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[6]  V. Oi.  Immunoglobulin producing hybrid cell lines , 1980 .

[7]  Chi-Wai Wong,et al.  Histone deacetylase associated with mSin3A mediates repression by the acute promyelocytic leukemia-associated PLZF protein , 1998, Oncogene.

[8]  A. Zelent,et al.  Reduced retinoic acid-sensitivities of nuclear receptor corepressor binding to PML- and PLZF-RARalpha underlie molecular pathogenesis and treatment of acute promyelocytic leukemia. , 1998, Blood.

[9]  Johnson M Liu,et al.  Abnormal microsomal detoxification implicated in Fanconi anemia group C by interaction of the FAC protein with NADPH cytochrome P450 reductase. , 1998, Blood.

[10]  A. D’Andrea,et al.  The Fanconi anemia polypeptide FACC is localized to the cytoplasm. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[11]  N. Aoki,et al.  Molecular cloning of the breakpoint for 3q27 translocation in B-cell lymphomas and leukemias. , 1994, Blood.

[12]  M. Buchwald,et al.  Molecular and Cellular Biology of Fanconi Anemia , 1992, The American journal of pediatric hematology/oncology.

[13]  G. Privé,et al.  Crystal structure of the BTB domain from PLZF. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Licht,et al.  The Promyelocytic Leukemia Zinc Finger Protein Affects Myeloid Cell Growth, Differentiation, and Apoptosis , 1998, Molecular and Cellular Biology.

[15]  G. Privé,et al.  The BTB domain, found primarily in zinc finger proteins, defines an evolutionarily conserved family that includes several developmentally regulated genes in Drosophila. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Rowley,et al.  Identification of a gene, MLL, that spans the breakpoint in 11q23 translocations associated with human leukemias. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Jonathan D. Licht,et al.  Deconstructing a Disease: RAR, Its Fusion Partners, and Their Roles in the Pathogenesis of Acute Promyelocytic Leukemia , 1999 .

[18]  M. Privalsky,et al.  Retinoid Isomers Differ in the Ability to Induce Release of SMRT Corepressor from Retinoic Acid Receptor-α* , 1999, The Journal of Biological Chemistry.

[19]  S. Baylin,et al.  p53 activates expression of HIC-1, a new candidate tumour suppressor gene on 17p13.3 , 1995, Nature Genetics.

[20]  Hans Joenje,et al.  Expression cloning of a cDNA for the major Fanconi anaemia gene, FAA , 1996, Nature Genetics.

[21]  R. Marmorstein,et al.  Overexpression, Purification, Characterization, and Crystallization of the BTB/POZ Domain from the PLZF Oncoprotein* , 1997, The Journal of Biological Chemistry.

[22]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[23]  P. Pandolfi,et al.  The BCL-6 proto-oncogene controls germinal-centre formation and Th2-type inflammation , 1997, Nature Genetics.

[24]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[25]  V. Michels,et al.  The Genetic Basis of Human Cancer , 1999 .

[26]  M. Buchwald,et al.  Fanconi anemia and novel strategies for therapy. , 1994, Blood.

[27]  R. Espinosa,et al.  Identification of the gene associated with the recurring chromosomal translocations t(3;14)(q27;q32) and t(3;22)(q27;q11) in B-cell lymphomas. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Q. Waisfisz,et al.  A physical complex of the Fanconi anemia proteins FANCG/XRCC9 and FANCA. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[29]  K. Kinzler,et al.  Cancer-susceptibility genes. Gatekeepers and caretakers. , 1997, Nature.

[30]  S. Elledge,et al.  The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit. , 1993, Genes & development.

[31]  D. Näf,et al.  Functional Activity of the Fanconi Anemia Protein FAA Requires FAC Binding and Nuclear Localization , 1998, Molecular and Cellular Biology.

[32]  D. Näf,et al.  Fanconi Anemia Proteins FANCA, FANCC, and FANCG/XRCC9 Interact in a Functional Nuclear Complex , 1999, Molecular and Cellular Biology.

[33]  C. Mathew,et al.  The Fanconi anemia group E gene, FANCE, maps to chromosome 6p. , 1999, American journal of human genetics.

[34]  L. Szekely,et al.  The Promyelocytic Leukemia Gene Product (PML) Forms Stable Complexes with the Retinoblastoma Protein , 1998, Molecular and Cellular Biology.

[35]  K. Schughart,et al.  Isolation and embryonic expression of the novel mouse gene Hic1, the homologue of HIC1, a candidate gene for the Miller-Dieker syndrome. , 1999, Human molecular genetics.

[36]  R. Gibson,et al.  A common mutation in the FACC gene causes Fanconi anaemia in Ashkenazi Jews , 1993, Nature Genetics.

[37]  E. Dmitrovsky,et al.  Leukemia translocation protein PLZF inhibits cell growth and expression of cyclin A , 1999, Oncogene.

[38]  C. Tribioli,et al.  Distinct interactions of PML-RARalpha and PLZF-RARalpha with co-repressors determine differential responses to RA in APL. , 1998, Nature genetics.

[39]  Albert B. Reynolds,et al.  The Catenin p120ctnInteracts with Kaiso, a Novel BTB/POZ Domain Zinc Finger Transcription Factor , 1999, Molecular and Cellular Biology.

[40]  H. Alder,et al.  The t(4;11) chromosome translocation of human acute leukemias fuses the ALL-1 gene, related to Drosophila trithorax, to the AF-4 gene , 1992, Cell.

[41]  P. Tiollais,et al.  The RAR alpha-PLZF chimera associated with Acute Promyelocytic Leukemia has retained a sequence-specific DNA-binding domain. , 1997, Oncogene.

[42]  M. Buchwald,et al.  Evidence for at least eight Fanconi anemia genes. , 1997, American journal of human genetics.

[43]  E. Craig,et al.  Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. , 1996, Genetics.

[44]  R. Moses,et al.  The Ashkenazi Jewish Fanconi anemia mutation: Incidence among patients and carrier frequency in the at‐risk population , 1994, Human mutation.

[45]  M. Privalsky,et al.  Components of the SMRT Corepressor Complex Exhibit Distinctive Interactions with the POZ Domain Oncoproteins PLZF, PLZF-RARα, and BCL-6* , 1998, The Journal of Biological Chemistry.

[46]  A D Auerbach,et al.  Phenotypic consequences of mutations in the Fanconi anemia FAC gene: an International Fanconi Anemia Registry study. , 1997, Blood.

[47]  M. Heinrich,et al.  The Fanconi anemia group C gene product is located in both the nucleus and cytoplasm of human cells. , 1998, Blood.

[48]  S. Kajigaya,et al.  Molecular chaperone GRP94 binds to the Fanconi anemia group C protein and regulates its intracellular expression. , 1998, Blood.

[49]  A. Dejean,et al.  SMRT corepressor interacts with PLZF and with the PML-retinoic acid receptor alpha (RARalpha) and PLZF-RARalpha oncoproteins associated with acute promyelocytic leukemia. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[50]  W. R. Shannon,et al.  Cloning of cDNAs for Fanconi's anaemia by functional complementation , 1992, Nature.

[51]  C. Mathew,et al.  A patient-derived mutant form of the Fanconi anemia protein, FANCA, is defective in nuclear accumulation. , 1999, Experimental hematology.

[52]  D. Leprince,et al.  Novel BTB/POZ domain zinc-finger protein, LRF, is a potential target of the LAZ-3/BCL-6 oncogene , 1999, Oncogene.

[53]  Yonghong Xiao,et al.  Association of BRCA1 with Rad51 in Mitotic and Meiotic Cells , 1997, Cell.

[54]  H. Joenje,et al.  Sequence variations in the Fanconi anaemia gene, FAC: pathogenicity of 1806insA and R548X and recognition of D195V as a polymorphic variant , 1996, Human Genetics.

[55]  J. Licht,et al.  Amino-terminal protein-protein interaction motif (POZ-domain) is responsible for activities of the promyelocytic leukemia zinc finger-retinoic acid receptor-alpha fusion protein. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[56]  A. Zelent,et al.  Fusion between a novel Krüppel‐like zinc finger gene and the retinoic acid receptor‐alpha locus due to a variant t(11;17) translocation associated with acute promyelocytic leukaemia. , 1993, The EMBO journal.

[57]  R. Berger,et al.  Rearrangements of the retinoic acid receptor alpha and promyelocytic leukemia zinc finger genes resulting from t(11;17)(q23;q21) in a patient with acute promyelocytic leukemia. , 1993, The Journal of clinical investigation.

[58]  R. Gale,et al.  Hematologic Abnormalities in Fanconi Anemia: An International Fanconi Anemia Registry Study , 1994 .

[59]  A. D’Andrea,et al.  The Fanconi anemia polypeptide, FAC, binds to the cyclin-dependent kinase, cdc2. , 1997, Blood.

[60]  D. Leprince,et al.  The BTB/POZ domain targets the LAZ3/BCL6 oncoprotein to nuclear dots and mediates homomerisation in vivo. , 1995, Oncogene.

[61]  Signaling by tyrosine kinases negatively regulates the interaction between transcription factors and SMRT (silencing mediator of retinoic acid and thyroid hormone receptor) corepressor. , 1998, Molecular endocrinology.

[62]  N. Andrews,et al.  A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. , 1991, Nucleic acids research.

[63]  M. Privalsky,et al.  Transcriptional Repression by the SMRT-mSin3 Corepressor: Multiple Interactions, Multiple Mechanisms, and a Potential Role for TFIIB , 1998, Molecular and Cellular Biology.

[64]  J M Davies,et al.  Leukemia-associated retinoic acid receptor alpha fusion partners, PML and PLZF, heterodimerize and colocalize to nuclear bodies. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[65]  K Offit,et al.  Alterations of a zinc finger-encoding gene, BCL-6, in diffuse large-cell lymphoma. , 1993, Science.

[66]  Hans Joenje,et al.  The Fanconi anaemia group G gene FANCG is identical with XRCC9 , 1998, Nature Genetics.

[67]  G. Fanconi.  Familial constitutional panmyelocytopathy, Fanconi's anemia (F.A.). I. Clinical aspects. , 1967, Seminars in hematology.

[68]  S. Latt,et al.  Flow cytometric characterization of the response of Fanconi's anemia cells to mitomycin C treatment. , 2005, Cytometry.

[69]  J. Licht,et al.  Reduced and altered DNA-binding and transcriptional properties of the PLZF-retinoic acid receptor-alpha chimera generated in t(11;17)-associated acute promyelocytic leukemia. , 1996, Oncogene.

[70]  L. Staudt,et al.  Control of inflammation, cytokine expression, and germinal center formation by BCL-6. , 1997, Science.

[71]  M. Buchwald.  Complementation groups: one or more per gene? , 1995, Nature Genetics.

[72]  M. Buchwald,et al.  Is Fanconi anemia caused by a defect in the processing of DNA damage? , 1998, Mutation research.

[73]  G. Evans,et al.  A trithorax–like gene is interrupted by chromosome 11q23 translocations in acute leukaemias , 1992, Nature Genetics.

[74]  A. Zelenetz,et al.  Clinical and molecular characterization of a rare syndrome of acute promyelocytic leukemia associated with translocation (11;17). , 1995, Blood.

[75]  D. Leprince,et al.  The BTB/POZ domain: a new protein-protein interaction motif common to DNA- and actin-binding proteins. , 1995, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[76]  R. Treisman,et al.  The POZ domain: a conserved protein-protein interaction motif. , 1994, Genes & development.

[77]  Michael L. Cleary,et al.  Involvement of a homolog of Drosophila trithorax by 11q23 chromosomal translocations in acute leukemias , 1992, Cell.

[78]  R. Krumlauf,et al.  Leukemia translocation gene, PLZF, is expressed with a speckled nuclear pattern in early hematopoietic progenitors. , 1995, Blood.