title: KPNA3-knockdown eliminates the second heat shock protein peak associated with the heat shock response of male silkworm pupae (Bombyx mori) by reducing heat shock factor transport into the nucleus.
abstract: In this study, we investigated the role of karyopherin alpha 3 in the heat shock response in male silkworm pupae. Karyopherin alpha recognizes the classical nuclear location sequence on proteins and transports them into the nucleus by forming a trimetric complex with karyopherin beta. Three predicted karyopherin alphas (KPNA1, KPNA2 and KPNA3) have been identified from the silkworm Bombyx mori. Pull-down assay result showed that KPNA3 can pull down heat shock transcription factor (HSF) from proteins extracted from tissues using non-denature lysis buffer. After 45 °C heat shock on male B. mori pupae for 30 min, we identified two heat shock protein (HSP) mRNA expression peaks correlating with HSP19.9, HSP20.4 and HSP25.4 at 4 h (peak 1) and 24 h (peak 2). The second peak was eliminated after knockdown of KPNA3. Similar results were obtained following knockdown of HSF, which is the trans-activating factor of heat shock. However, KPNA3 knockdown was not accompanied by the decreased HSF protein levels at 24 h after heat shock which were observed following HSF knockdown. We also expressed recombinant protein GST-KPNA3 and His-HSF in Escherichia coli to perform GST pull-down assay and the result confirmed the interaction between KPNA3 and HSF. We concluded that KPNA3 knockdown eliminates the second heat shock protein peak in the heat shock response of male silkworm pupae by reducing HSF transport into the nucleus.
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China; College of Life Sciences, Huaibei Normal University, Huaibei, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China.
authors.forename: Guoqing
authors.initials: G
authors.lastname: Wei
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China.
authors.forename: Cen
authors.initials: C
authors.lastname: Qian
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China.
authors.forename: Kedong
authors.initials: K
authors.lastname: Li
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China.
authors.forename: Congfen
authors.initials: C
authors.lastname: Zhang
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China.
authors.forename: Lishang
authors.initials: L
authors.lastname: Dai
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China.
authors.forename: Dongran
authors.initials: D
authors.lastname: Liu
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China. Electronic address: zhubaojian@ahau.edu.cn.
authors.forename: Baojian
authors.initials: B
authors.lastname: Zhu
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China. Electronic address: cyschx@163.com.
authors.forename: Chaoliang
authors.initials: C
authors.lastname: Liu
mesh_terms: D000818:Animals; D012831:Bombyx / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018869:Heat-Shock Response; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D028901:alpha Karyopherins / Q000235:genetics* / Q000378:metabolism
chemical_list: D006360:Heat-Shock Proteins; D019476:Insect Proteins; D014157:Transcription Factors; D028901:alpha Karyopherins
keywords: Bombyx mori; Heat shock response; Heat shock transcription factor; Karyopherin alpha 3; RNA interference