Translocation of N-WASP by nuclear localization and export signals into the nucleus modulates expression of HSP90.
N-WASP regulates the actin cytoskeleton through activation of the Arp2/3 complex. N-WASP localizes at the cell periphery, where it controls actin polymerization downstream of signal molecules such as adapter proteins, Cdc42, Src family kinases, and phosphoinositides. N-WASP also localizes in the nucleus; however, the role of N-WASP in the nucleus is unclear. Here, we show that localization of N-WASP is controlled through phosphorylation by Src family kinases in which phosphorylated N-WASP is exported from the nucleus in a nuclear export signal (NES) and leptomycin B-dependent manner. N-WASP had nuclear localization signal (NLS) at its basic region and NES close to the phosphorylation site by Src family kinases, indicating that phosphorylation controls the accessibility to the NES through conformational changes. Increased levels of unphosphorylated N-WASP in the nucleus suppressed expression of HSP90 and transcription from a heat shock element (HSE). N-WASP bound heat shock transcription factor (HSTF) and enhanced the HSTF association with HSE. In addition, nuclear N-WASP was present in the protein complex that associates with HSE, suggesting that N-WASP participates in suppression of HSP90 transcription. Increased levels of unphosphorylated N-WASP also decreased the activities of Src family kinases in cells but not in experiments in vitro with pure N-WASP and Fyn. Because HSP90 is essential for the activities of Src family kinases, these results suggest that localization of N-WASP modulates Src kinase activity by regulating HSP90 expression.
Suetsugu
Shiro
S
Department of Biochemistry, Institute of Medical Science, University of Tokyo, Japan.
Takenawa
Tadaomi
T
D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D002460:Cell Line; D015536:Down-Regulation; D005231:Fatty Acids, Unsaturated; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D019913:Nuclear Localization Signals / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation; D020218:Response Elements; D014162:Transfection; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; D019061:src-Family Kinases / Q000378:metabolism
D005231:Fatty Acids, Unsaturated; D018841:HSP90 Heat-Shock Proteins; D009419:Nerve Tissue Proteins; D019913:Nuclear Localization Signals; C495771:WASL protein, human; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; D019061:src-Family Kinases; C038753:leptomycin B