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PMID: 10866674 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The docking protein HEF1 is an apoptotic mediator at focal adhesion sites.

Molecular and cellular biology ·Vol. 20 ·No. 14 ·2000-07-00 ·Pages 5184-95

Law SF, O'Neill GM, Fashena SJ, Einarson MB, Golemis EA

Abstract

HEF1 (human enhancer of filamentation 1) is a member of a docking protein family that includes p130(Cas) and Efs. Through assembly of multiple protein interactions at focal adhesion sites, these proteins activate signaling cascades in response to integrin receptor binding of the extracellular matrix. The HEF1 protein is cell cycle regulated, with full-length forms cleaved in mitosis at a caspase consensus site to generate an amino-terminal 55-kDa form that localizes to the mitotic spindle. The identification of a caspase cleavage site in HEF1 led us to investigate whether HEF1 belongs to a select group of caspase substrates cleaved in apoptosis to promote the morphological changes characteristic of programmed cell death. Significantly, inducing expression of HEF1 in MCF-7 or HeLa cells causes extensive apoptosis, as assessed by multiple criteria. Endogenous HEF1 is cleaved into 65- and 55-kDa fragments and a newly detected 28-kDa form in response to the induction of apoptosis, paralleling cleavage of poly(ADP-ribose) polymerase and focal adhesion kinase (FAK); the death-promoting activity of over-expressed HEF1 is associated with production of the 28-kDa form. While the generation of the cleaved HEF1 forms is caspase dependent, the accumulation of HEF1 forms is further regulated by the proteasome, as the proteasome inhibitors N-acetyl-L-leucinyl-L-leucinyl-L-norleucinyl and lactacystin enhance their stability. Finally, the induction of HEF1 expression also increases Jun N-terminal protein kinase (JNK) activation, and activated JNK colocalizes with HEF1, implicating this pathway in HEF1 action. Based on these results, we propose that dysregulation of HEF1 and its family members along with FAK may signal the destruction of focal adhesion sites and regulate the onset of apoptosis.

MeSH Terms
Acetylcysteine/analogs & derivatives,pharmacology Adaptor Proteins, Signal Transducing Apoptosis/drug effects,physiology Caspase 3 Caspase 7 Caspase Inhibitors Caspases/metabolism Cell Death/physiology Crk-Associated Substrate Protein Cysteine Endopeptidases/metabolism Cysteine Proteinase Inhibitors/pharmacology Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases HeLa Cells Humans JNK Mitogen-Activated Protein Kinases Leupeptins/pharmacology Mitogen-Activated Protein Kinases/metabolism Mitosis Multienzyme Complexes/metabolism Mutation Oligopeptides/pharmacology Peptide Fragments/genetics,metabolism Phosphoproteins/drug effects,genetics,metabolism Poly(ADP-ribose) Polymerases/metabolism Proteasome Endopeptidase Complex Protein Isoforms/metabolism Protein-Tyrosine Kinases/metabolism Proteins Retinoblastoma-Like Protein p130 Signal Transduction Tumor Cells, Cultured Tumor Necrosis Factor-alpha/pharmacology
Chemicals
Adaptor Proteins, Signal Transducing BCAR1 protein, human Caspase Inhibitors Crk-Associated Substrate Protein Cysteine Proteinase Inhibitors Leupeptins Multienzyme Complexes NEDD9 protein, human Oligopeptides Peptide Fragments Phosphoproteins Protein Isoforms Proteins Retinoblastoma-Like Protein p130 Tumor Necrosis Factor-alpha benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone acetylleucyl-leucyl-norleucinal lactacystin Poly(ADP-ribose) Polymerases Protein-Tyrosine Kinases Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases PTK2 protein, human JNK Mitogen-Activated Protein Kinases Mitogen-Activated Protein Kinases CASP3 protein, human CASP7 protein, human Caspase 3 Caspase 7 Caspases Cysteine Endopeptidases Proteasome Endopeptidase Complex Acetylcysteine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Law S F
Division of Basic Science, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA.
O'Neill G M
Fashena S J
Einarson M B
Golemis E A
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.20.14.5184-5195.2000
Published
2000-07-00
Pages
5184-95
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC85967
Subset
IM
Grants
NCI NIH HHS · R01 CA63366 · United States
NIGMS NIH HHS · F32 GM018223 · United States
NCI NIH HHS · R01 CA063366 · United States
NIGMS NIH HHS · F32 GM18223 · United States
NCI NIH HHS · P30 CA006927 · United States
NCI NIH HHS · CA-06927 · United States
NCI NIH HHS · T32 CA009035 · United States
External Links
PubMed source DOI: 10.1128/MCB.20.14.5184-5195.2000 PMC Full Text View journal details
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