Expression of MARCKS effector domain mutants alters phospholipase D activity and cytoskeletal morphology of SK-N-MC neuroblastoma cells

Sherry C. Morash, Donna Douglas, Christopher R. McMaster, Harold W. Cook, David M. Byers

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6 Citas (Scopus)

Resumen

Stable overexpression of myristoylated alanine-rich C-kinase substrate (MARCKS) is known to enhance phorbol ester stimulation of phospholipase D (PLD) activity and protein kinase Cα (PKCα) levels in SK-N-MC neuroblastoma cells. In contrast, expression of MARCKS mutants (S152A or S156A) lacking key PKC phosphorylation sites within the central basic effector domain (ED) had no significant effect on PLD activity or PKCα levels relative to vector control cells. Like control cells, those expressing wild type MARCKS were elongated and possessed longitudinally oriented stress fibers, although these cells were more prone to detach from the substratum and undergo cell death upon phorbol ester treatment. However, cells expressing MARCKS ED mutants were irregularly shaped and stress fibers were either shorter or less abundant, and cell adhesion and viability were not affected. These results suggest that intact phosphorylation sites within the MARCKS ED are required for PLD activation and influence both membrane-cytoskeletal organization and cell viability.

Idioma originalEnglish
Páginas (desde-hasta)1353-1364
Número de páginas12
PublicaciónNeurochemical Research
Volumen30
N.º11
DOI
EstadoPublished - nov. 2005

Nota bibliográfica

Funding Information:
This work was supported by operating grants from the Canadian Institutes for Health Research (DMB, HWC, and CRM), the Lockwood Foundation (SCM), and a Canada Research Chairs award (CRM). The excellent technical assistance of Susan Powell, Robert Zwicker and Gladys Keddy is gratefully acknowledged. The authors thank Dr. Michael Frohman, State University of New York, Stony Brook for the generous gifts of the PLD1 antibody and HA-tagged hPLD1 and mPLD2 constructs, and Dr. Sylvain Bourgoin, Centre de Recherche du CHUL, Quebec, Canada for the PLD2 antibody.

ASJC Scopus Subject Areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't

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