title: Inhibition of cellular proliferation by the Wilms tumor suppressor WT1 requires association with the inducible chaperone Hsp70.
abstract: The Wilms tumor suppressor WT1 encodes a zinc finger transcription factor that is expressed in glomerular podocytes during a narrow window in kidney development. By immunoprecipitation and protein microsequencing analysis, we have identified a major cellular protein associated with endogenous WT1 to be the inducible chaperone Hsp70. WT1 and Hsp70 are physically associated in embryonic rat kidney cells, in primary Wilms tumor specimens and in cultured cells with inducible expression of WT1. Colocalization of WT1 and Hsp70 is evident within podocytes of the developing kidney, and Hsp70 is recruited to the characteristic subnuclear clusters that contain WT1. The amino-terminal transactivation domain of WT1 is required for binding to Hsp70, and expression of that domain itself is sufficient to induce expression of Hsp70 through the heat shock element (HSE). Substitution of a heterologous Hsp70-binding domain derived from human DNAJ is sufficient to restore the functional properties of a WT1 protein with an amino-terminal deletion, an effect that is abrogated by a point mutation in DNAJ that reduces binding to Hsp70. These observations indicate that Hsp70 is an important cofactor for the function of WT1, and suggest a potential role for this chaperone during kidney differentiation.
authors.affiliation: Massachusetts General Hospital Cancer Center, Massachusetts General Hospital (MGH) and Harvard Medical School, Charlestown, Massachusetts 02129, USA.
authors.forename: S
authors.initials: S
authors.lastname: Maheswaran
authors.forename: C
authors.initials: C
authors.lastname: Englert
authors.forename: G
authors.initials: G
authors.lastname: Zheng
authors.forename: S B
authors.initials: SB
authors.lastname: Lee
authors.forename: J
authors.initials: J
authors.lastname: Wong
authors.forename: D P
authors.initials: DP
authors.lastname: Harkin
authors.forename: J
authors.initials: J
authors.lastname: Bean
authors.forename: R
authors.initials: R
authors.lastname: Ezzell
authors.forename: A J
authors.initials: AJ
authors.lastname: Garvin
authors.forename: R T
authors.initials: RT
authors.lastname: McCluskey
authors.forename: J A
authors.initials: JA
authors.lastname: DeCaprio
authors.forename: D A
authors.initials: DA
authors.lastname: Haber
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002455:Cell Division; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005786:Gene Expression Regulation; D016162:Genes, Wilms Tumor; D006131:Growth Inhibitors / Q000378:metabolism*; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000196:embryology / Q000378:metabolism; D007678:Kidney Glomerulus / Q000196:embryology / Q000378:metabolism; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D051381:Rats; D014157:Transcription Factors / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D025721:WT1 Proteins; D016335:Zinc Fingers
chemical_list: D004268:DNA-Binding Proteins; D006131:Growth Inhibitors; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D014157:Transcription Factors; D025721:WT1 Proteins