Molecular and Cellular Biochemistry

, Volume 366, Issue 1–2, pp 175–182 | Cite as

Sar1 translocation onto the ER-membrane for vesicle budding has different pathways for promotion and suppression of ER-to-Golgi transport mediated through H89-sensitive kinase and ER-resident G protein

  • Hiroshi Nakagawa
  • Masakazu Ishizaki
  • Shuichi Miyazaki
  • Takuto Abe
  • Kazuhiko Nishimura
  • Masayuki Komori
  • Saburo MatsuoEmail author


ER-to-Golgi protein transport involves transport vesicles of which formation is initiated by assembly of Sar1. The assembly of Sar1 is suppressed by protein kinase inhibitor H89, suggesting that ER-to-Golgi transport is regulated progressively by H89 sensitive kinase. ER-resident Gi2 protein suppresses vesicle formation with inhibition of Sar1 assembly. This study examined whether these promotion and suppression of vesicle transport share the same signal pathway, by examining the effects of Gi/o protein activator mastoparan 7 (Mp-7) and H89 on Sar1 and Sec23 recruitment onto microsomes. In a cell-free system for Sar1 translocation assay, GTPγS addition induced the translocation of Sar1 onto microsomes. Mp-7 and H89 decreased the Sar1 translocation. Double treatment of Mp-7 and H89 strongly decreased Sar1 translocation. In single and double treatments, however, Gi/o protein inactivator pertussis toxin (IAP) partially restored the suppressive effect of Mp-7, but had not any effect on H89-induced effect. Then, the assembly of Sec23 onto the microsome was also increased by the addition of GTPγS. Sec23 translocation was decreased by Mp-7 and/or H89 treatment and recovered by IAP pretreatment except for H89 single treatment, similarly to Sar1 translocation in each treatment. Inhibitory effects of H89 and Mp-7on ER-to-Golgi vesicle transport by H89 or Mp-7 were also confirmed in a cell culture system by BFA-dispersion and BFA-reconstruction experiments. These findings indicate that promotion and suppression of ER-to-Golgi vesicle transport are modulated through separate signal pathways.


ER-to-Golgi transport Sar1 Coat assembly H89 sensitive kinase ER-resident Gi protein 


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Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Hiroshi Nakagawa
    • 1
  • Masakazu Ishizaki
    • 1
  • Shuichi Miyazaki
    • 2
  • Takuto Abe
    • 1
  • Kazuhiko Nishimura
    • 1
  • Masayuki Komori
    • 3
  • Saburo Matsuo
    • 1
    Email author
  1. 1.Laboratory of Toxicology, Course of Veterinary Science, Graduate School of Life and Environmental BiosciencesOsaka Prefecture UniversityIzumisanoJapan
  2. 2.The Center for Advanced Research, Graduate School of Medical SciencesToho University of MedicineTokyoJapan
  3. 3.Laboratory of Cellular and Molecular Biology, Course of Veterinary Science, Graduate School of Life and Environmental BiosciencesOsaka Prefecture UniversityIzumisanoJapan

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