Site-directed mutation of nm23-H1 - Mutations lacking motility suppressive capacity upon transfection are deficient in histidine-dependent protein phosphotransferase pathways in vitro

被引：124

作者：

Freije, JMP ^{[1
]}

Blay, P ^{[1
]}

MacDonald, NJ ^{[1
]}

Manrow, RE ^{[1
]}

Steeg, PS ^{[1
]}

机构：

[1] NCI, WOMENS CANC SECT, PATHOL LAB, DIV CLIN SCI, NIH, BETHESDA, MD 20892 USA

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 1997年 / 272卷 / 09期

关键词：

D O I：

10.1074/jbc.272.9.5525

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：

We previously compared the structure and motility suppressive capacity of nm23-H1 by transfection of wild type and site-directed mutant forms into breast carcinoma cells, Wild type nm23-H1 and an nm23-H1(S44A) (serine 44 to alanine) mutant suppressed motility, whereas the nm23-H1(P96S), nm23-H1(S120G), and to a lesser extent, nm23-H1(S120A) mutant forms failed to do so, In the present study wild type and mutant recombinant Nm23-H1 proteins have been produced, purified, and assayed for phosphorylation and phosphotransfer activities, We report the first association of Nm23-H1 mutations lacking motility suppressive capacity with decreased in vitro activity in histidine-dependent protein phosphotransferase assays. Nm23-H1(P96S), a Drosophila developmental mutation homolog, exhibited normal autophosphorylation and nucleoside-diphosphate kinase (NDPK) characteristics but deficient phosphotransfer activity in three histidine protein kinase assays, using succinic thiokinase, Nm23-H2, and GST-Nm23-H1 as substrates. Nm23-H1(S120G), found in advanced human neuroblastomas, exhibited deficient activity in several histidine-dependent protein phosphotransfer reactions, including histidine autophosphorylation, downstream phosphorylation on serines, and slightly decreased histidine protein kinase activity; significant NDPK activity was observed. The Nm23-H1(S120A) mutant was deficient in only histidine-dependent serine autophosphorylation. Nm23-H1 and Nm23-H1(S44A) exhibited normal activity in all assays conducted. Based on this correlation, we hypothesize that a histidine-dependent protein phosphotransfer activity of Nm23-H1 may be responsible for its biological suppressive effects.

引用

页码：5525 / 5532

页数：8

共 62 条

[1] PROTEIN HISTIDINE KINASES AND SIGNAL-TRANSDUCTION IN PROKARYOTES AND EUKARYOTES [J].

ALEX, LA ;

SIMON, MI .

TRENDS IN GENETICS, 1994, 10 (04) :133-138

[2]

BABA H, 1995, CANCER RES, V55, P1977

[3]

BERBERICH S, 1995, ONCOGENE, P2343

[4] TRANSPHOSPHORYLATION BETWEEN NUCLEOSIDE POLYPHOSPHATES [J].

BERG, P ;

JOKLIK, WK .

NATURE, 1953, 172 (4387) :1008-1009

[5] ANALYSIS OF THE LETHAL INTERACTION BETWEEN THE PRUNE AND KILLER OF PRUNE MUTATIONS OF DROSOPHILA [J].

BIGGS, J ;

TRIPOULAS, N ;

HERSPERGER, E ;

DEAROLF, C ;

SHEARN, A .

GENES & DEVELOPMENT, 1988, 2 (10) :1333-1343

[6] A DROSOPHILA GENE THAT IS HOMOLOGOUS TO A MAMMALIAN GENE ASSOCIATED WITH TUMOR-METASTASIS CODES FOR A NUCLEOSIDE DIPHOSPHATE KINASE [J].

BIGGS, J ;

HERSPERGER, E ;

STEEG, PS ;

LIOTTA, LA ;

SHEARN, A .

CELL, 1990, 63 (05) :933-940

[7] AUTOPHOSPHORYLATION OF NUCLEOSIDE DIPHOSPHATE KINASE ON NON-HISTIDINE RESIDUES [J].

BOMINAAR, AA ;

TEPPER, AD ;

VERON, M .

FEBS LETTERS, 1994, 353 (01) :5-8

[8] NM23-H1 MUTATION IN NEUROBLASTOMA [J].

CHANG, CL ;

ZHU, XX ;

THORAVAL, DH ;

UNGAR, D ;

RAWWAS, J ;

HORA, N ;

STRAHLER, JR ;

HANASH, SM ;

RADANY, E .

NATURE, 1994, 370 (6488) :335-336

[9] CRYSTAL-STRUCTURE OF THE AWD NUCLEOTIDE DIPHOSPHATE KINASE FROM DROSOPHILA [J].

CHIADMI, M ;

MORERA, S ;

LASCU, I ;

DUMAS, C ;

LEBRAS, G ;

VERON, M ;

JANIN, J .

STRUCTURE, 1993, 1 (04) :283-293

[10] HISTIDINE PHOSPHORYLATION OF P-SELECTIN UPON STIMULATION OF HUMAN PLATELETS - A NOVEL PATHWAY FOR ACTIVATION-DEPENDENT SIGNAL-TRANSDUCTION [J].

CROVELLO, CS ;

FURIE, BC ;

FURIE, B .

CELL, 1995, 82 (02) :279-286

← 1 2 3 4 5 6 7 →