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Regulation of vesicle transport and cell motility by Golgi-localized Dbs

Ethan R Fitzpatrick1, Tinghui Hu, Bryan T Ciccarelli

  • 1a Department of Microbiology and Molecular Genetics; The New Jersey Medical School-Cancer Center; Rutgers Biomedical and Health Sciences ; Newark , NJ USA.

Small Gtpases
|December 9, 2014
PubMed

Insights

Dbs-130, a protein involved in cell motility and osteoarthritis risk, regulates Golgi function and vesicle transport. Inhibiting Dbs-130 impairs cell migration by affecting Golgi reorientation during directed movement.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • DBS/MCF2L is a risk locus for osteoarthritis, encoding a guanine nucleotide exchange factor (Dbs).
  • Dbs regulates normal and tumor cell motility.
  • Endogenous Dbs exists as two isoforms: Dbs-130 (Golgi-localized) and Dbs-80 (ER-localized).

Purpose of the Study:

  • To investigate the function of Dbs isoforms, particularly Dbs-130, in cellular processes.
  • To determine the role of Dbs-130 in vesicle transport and cell migration.
  • To explore the therapeutic potential of Dbs inhibition.

Main Methods:

  • Utilized an inhibitor targeting the RhoGEF domain of Dbs.
  • Investigated inhibitor localization and interaction with Dbs-130.
  • Assessed effects of Dbs-130 inhibition on activated Cdc42 levels, Golgi morphology, and Brefeldin A sensitivity.
  • Analyzed protein transport from ER to Golgi and Golgi to plasma membrane.
  • Performed transwell and wound healing assays in MDA-MB-231 breast tumor cells.
  • Examined Golgi reorientation and microtubule cytoskeleton organization.

Main Results:

  • The Dbs inhibitor specifically interacts with Dbs-130 at the Golgi.
  • Inhibition of Dbs-130 reduced activated Cdc42, enlarged the Golgi, and conferred resistance to Brefeldin A.
  • ER-to-Golgi transport remained normal, but Golgi-to-plasma membrane transport was impaired.
  • Dbs-130 inhibition in breast cancer cells reduced motility without affecting microtubule organization.
  • Reduced motility correlated with a failure to reorient the Golgi toward the leading edge.

Conclusions:

  • Dbs-130 plays a critical role in vesicle transport and the secretory pathway.
  • Dbs-130 is essential for Golgi reorientation and cell polarization during directed migration.
  • Targeting Dbs-130 may offer a strategy to limit tumor cell motility.
Keywords:
BFA, Brefeldin ACOP1, coat proteinDAPI, 4’, 6-diamidino-2-phenylindoleDH, Dbl homologyDbsDbs, Dbl's big sisterECL, electrochemiluminescenceER, endoplasmic reticulumFACS, fluorescence-activated cell sortingFGD1, faciogenital dysplasia 1 proteinGEF, guanine nucleotide exchange factorGFP, green fluorescent proteinHA, hemagglutininHM, homogenization mediumMCF2LMTOC, microtubule organizing centerNF-1, neurofibromatosis type 1PAK3, p21 protein-activated kinase 3PBS, phosphate buffered salinePH, pleckstrin homologyRhoSH3, Src homology 3VSVG, vesicular stomatitis virus-GWGA, wheat germ agglutininbreast cancercell motilitygolgi complexguanine nucleotide exchange factorosteoarthritisrtPCR, real-time polymerase chain reactionsiRNA, small inhibitory RNA

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