EARLY ACTIVATION OF ENDOGENOUS PP60SRC KINASE-ACTIVITY DURING NEURONAL DIFFERENTIATION OF CULTURED HUMAN NEUROBLASTOMA-CELLS

被引:95
作者
BJELFMAN, C
MEYERSON, G
CARTWRIGHT, CA
MELLSTROM, K
HAMMERLING, U
PAHLMAN, S
机构
[1] UNIV UPPSALA HOSP,DEPT PATHOL,UPPSALA,SWEDEN
[2] SALK INST BIOL STUDIES,MOLEC BIOL & VIROL LAB,SAN DIEGO,CA 92138
关键词
D O I
10.1128/MCB.10.1.361
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The proto-oncogene product pp60(c-src) is a tyrosine-specific kinase with a still unresolved cellular function. High levels of pp60(c-src) in neurons and the existence of a neuronal pp60(c-src) variant, pp60(c-srcN), suggest participation in the progress or maintenance of the differentiated phenotype of neurons. We have previously reported that phorbol esters, e.g., 12-O-tetradecanoylphorbol-13-acetate (TPA), stimulate human SH-SY5Y neuroblastoma cells to neuronal differentiation, as monitored by morphological, biochemical, and functional differentiation markers. In this report, we describe activation of the pp60(src) (pp60(c-src) and pp60(c-srcN)) kinase activity observed at 6 h after induction of SH-SY5Y cells with TPA. This phenomenon coincides in time with neurite outgrowth, formation of growth cone-like structures, and an increase of GAP43 mRNA expression, which are the earliest indications of neuronal differentiation in these cells. The highest specific src kinase activity (a three- to fourfold increase 4 days after induction) was noted in cells treated with 16 nM TPA; this concentration is optimal for development of the TPA-induced neuronal phenotype. During differentiation, there was no alteration in the 1:1 ratio of pp60(c-src) to pp60(c-srcN) found in untreated SH-SY5Y cells. V8 protease and trypsin phosphopeptide mapping of pp60(src) from in vivo 32P-labeled cells showed that the overall phosphorylation of pp60(src) was higher in differentiated than in untreated cells, mainly because of an intense serine 12 phosphorylation. Tyrosine 416 phosphorylation was not detectable in either cell type, and no change during differentiation in tyrosine 527 phosphorylation was observed.
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页码:361 / 370
页数:10
相关论文
共 66 条
[1]   MUSCARINIC RECEPTORS IN HUMAN SH-SY5Y NEUROBLASTOMA CELL-LINE - REGULATION BY PHORBOL ESTER AND RETINOIC ACID-INDUCED DIFFERENTIATION [J].
ADEM, A ;
MATTSSON, MEK ;
NORDBERG, A ;
PAHLMAN, S .
DEVELOPMENTAL BRAIN RESEARCH, 1987, 33 (02) :235-242
[2]   FUNCTIONAL-DIFFERENTIATION OF A HUMAN GANGLION-CELL DERIVED NEURO-BLASTOMA CELL-LINE SH-SY5Y INDUCED BY A PHORBOL ESTER (TPA) [J].
AKERMAN, KEO ;
SCOTT, IG ;
ANDERSSON, LC .
NEUROCHEMISTRY INTERNATIONAL, 1984, 6 (01) :77-80
[3]   RAPID AND REVERSIBLE REDUCTION OF JUNCTIONAL PERMEABILITY IN CELLS INFECTED WITH A TEMPERATURE-SENSITIVE MUTANT OF AVIAN-SARCOMA VIRUS [J].
ATKINSON, MM ;
MENKO, AS ;
JOHNSON, RG ;
SHEPPARD, JR ;
SHERIDAN, JD .
JOURNAL OF CELL BIOLOGY, 1981, 91 (02) :573-578
[4]   THE CELLULAR SRC GENE-PRODUCT REGULATES JUNCTIONAL CELL-TO-CELL COMMUNICATION [J].
AZARNIA, R ;
REDDY, S ;
KMIECIK, TE ;
SHALLOWAY, D ;
LOEWENSTEIN, WR .
SCIENCE, 1988, 239 (4838) :398-401
[5]   PRIMARY STRUCTURE AND TRANSCRIPTIONAL REGULATION OF GAP-43, A PROTEIN ASSOCIATED WITH NERVE GROWTH [J].
BASI, GS ;
JACOBSON, RD ;
VIRAG, I ;
SCHILLING, J ;
SKENE, JHP .
CELL, 1987, 49 (06) :785-791
[6]   CHARACTERIZATION OF ROUS-SARCOMA VIRUS SRC GENE PRODUCTS SYNTHESIZED INVITRO [J].
BEEMON, K ;
HUNTER, T .
JOURNAL OF VIROLOGY, 1978, 28 (02) :551-566
[7]  
BIEDLER JL, 1973, CANCER RES, V33, P2643
[8]  
BIEDLER JL, 1978, CANCER RES, V38, P3751
[9]   INCREASED PP60C-SRC TYROSYL KINASE-ACTIVITY IN HUMAN NEUROBLASTOMAS IS ASSOCIATED WITH AMINO-TERMINAL TYROSINE PHOSPHORYLATION OF THE SRC GENE-PRODUCT [J].
BOLEN, JB ;
ROSEN, N ;
ISRAEL, MA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (21) :7275-7279
[10]   CHARACTERIZATION OF THE ALTERED FORM OF THE C-SRC GENE-PRODUCT IN NEURONAL CELLS [J].
BRUGGE, J ;
COTTON, P ;
LUSTIG, A ;
YONEMOTO, W ;
LIPSICH, L ;
COUSSENS, P ;
BARRETT, JN ;
NONNER, D ;
KEANE, RW .
GENES & DEVELOPMENT, 1987, 1 (03) :287-296