Different localization of Hsp105 family proteins in mammalian cells.
Hsp105alpha and Hsp105beta of the HSP105 family are alternatively spliced products derived from an hsp 105 gene transcript. Hsp105alpha is constitutively expressed and also induced by various stress, whereas Hsp105beta, lacking 44 amino acids from Hsp105alpha, is specifically expressed during mild heat shock. Although Hsp105alpha is shown to localize in the cytoplasm of mammalian cells, cellular localization of Hsp105beta is not known. In this study, we showed that Hsp105beta localized in the nucleus of cells in contrast to cytoplasmic Hsp105alpha, suggesting that these proteins function in different cellular compartments of cells. Using deletion and substitution mutants of Hsp105alpha and Hsp105beta, we revealed that these proteins had a functional nuclear localization signal (NLS) and a nuclear export signal (NES). Furthermore, Hsp105alpha accumulated in the nucleus of cells when treated with leptomycin B, a specific inhibitor of NES-dependent nuclear export. siRNA for importin beta, an essential component for NLS-dependent nuclear transport, inhibited the nuclear localization of Hsp105beta. Furthermore, the 44 amino acids sequence found in Hsp105alpha but not in Hsp105beta suppressed the NLS activity. Thus, the different localization of Hsp105alpha and Hsp105beta is suggested to be due to the suppressed NLS activity in Hsp105alpha.
Saito
Youhei
Y
Department of Biochemistry, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
Yamagishi
Nobuyuki
N
Hatayama
Takumi
T
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002467:Cell Nucleus / Q000737:chemistry* / Q000378:metabolism; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000737:chemistry* / Q000378:metabolism; D050957:HSP110 Heat-Shock Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing
D050957:HSP110 Heat-Shock Proteins; C490783:Hsp105 protein, mouse; D019913:Nuclear Localization Signals; D008156:Luciferases