PHOSPHORYLATION OF CYCLIC ADENOSINE-3',5'-MONOPHOSPHATE (CAMP) RESPONSE ELEMENT-BINDING PROTEIN ISOFORMS BY THE CAMP-DEPENDENT PROTEIN-KINASE

被引:32
作者
SUN, PQ
SCHODERBEK, WE
MAURER, RA
机构
关键词
D O I
10.1210/me.6.11.1858
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The cAMP response element-binding protein (CREB) mediates transcriptional activation of genes in response to the cAMP signal transduction pathway. There are two different isoforms of CREB, which are generated by alternative RNA splicing. There is evidence that the two isoforms may have different biological activities. As the longer isoform (CREB341) contains a potential phosphorylation site that is not present in the shorter isoform (CREB327), we examined the possible differential phosphorylation of the two CREB isoforms. Recombinant CREB was prepared and used as substrate for phosphorylation by the cAMP-dependent protein kinase in vitro. Phosphopeptide mapping and mutagenesis studies demonstrated that CREB341 contains two sites, serine 133 and serine 98, that can be phosphorylated in vitro by the catalytic subunit of the cAMP-dependent protein kinase. In contrast, CREB327 contains only a single phosphorylation site at serine 119 (equivalent position to serine 133 in CREB341). A kinase titration experiment demonstrated that serine 98 of CREB341 was phosphorylated only at relatively high concentrations of the cAMP-dependent protein kinase. Transient transfection studies were used to test for any possible function of the phosphorylation of serine 98 of CREB341. These studies used GAL4-CREB fusion proteins. We found that mutation of serine 98 to alanine (which would block phosphorylation) has little or no effect on the ability of the CREB fusion protein to activate transcription. These findings suggest that differences in the biological activity of the two CREB isoforms are probably not mediated by differential phosphorylation by the cAMP-dependent protein kinase.
引用
收藏
页码:1858 / 1866
页数:9
相关论文
共 38 条
[1]   INVITRO TRANSCRIPTION DIRECTED FROM THE SOMATOSTATIN PROMOTER IS DEPENDENT UPON A PURIFIED 43-KDA DNA-BINDING PROTEIN [J].
ANDRISANI, OM ;
ZHU, Z ;
POT, DA ;
DIXON, JE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (07) :2181-2185
[2]   IDENTIFICATION AND REGULATION OF THE CYSTIC-FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR-GENERATED CHLORIDE CHANNEL [J].
BERGER, HA ;
ANDERSON, MP ;
GREGORY, RJ ;
THOMPSON, S ;
HOWARD, PW ;
MAURER, RA ;
MULLIGAN, R ;
SMITH, AE ;
WELSH, MJ .
JOURNAL OF CLINICAL INVESTIGATION, 1991, 88 (04) :1422-1431
[3]   2 DISTINCT FORMS OF ACTIVE TRANSCRIPTION FACTOR CREB (CAMP RESPONSE ELEMENT BINDING-PROTEIN) [J].
BERKOWITZ, LA ;
GILMAN, MZ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (14) :5258-5262
[4]  
BOYLE WJ, 1991, METHOD ENZYMOL, V201, P110
[5]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[6]   CAMP RESPONSE ELEMENT-BINDING PROTEIN IS ACTIVATED BY CA2+/CALMODULIN-DEPENDENT AS WELL AS CAMP-DEPENDENT PROTEIN-KINASE [J].
DASH, PK ;
KARL, KA ;
COLICOS, MA ;
PRYWES, R ;
KANDEL, ER .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (11) :5061-5065
[7]   FIREFLY LUCIFERASE GENE - STRUCTURE AND EXPRESSION IN MAMMALIAN-CELLS [J].
DEWET, JR ;
WOOD, KV ;
DELUCA, M ;
HELINSKI, DR ;
SUBRAMANI, S .
MOLECULAR AND CELLULAR BIOLOGY, 1987, 7 (02) :725-737
[8]   BOTH THE BASIC REGION AND THE LEUCINE ZIPPER DOMAIN OF THE CYCLIC-AMP RESPONSE ELEMENT BINDING (CREB) PROTEIN ARE ESSENTIAL FOR TRANSCRIPTIONAL ACTIVATION [J].
DWARKI, VJ ;
MONTMINY, M ;
VERMA, IM .
EMBO JOURNAL, 1990, 9 (01) :225-232
[9]   CHARACTERIZATION OF MOTIFS WHICH ARE CRITICAL FOR ACTIVITY OF THE CYCLIC AMP-RESPONSIVE TRANSCRIPTION FACTOR CREB [J].
GONZALEZ, GA ;
MENZEL, P ;
LEONARD, J ;
FISCHER, WH ;
MONTMINY, MR .
MOLECULAR AND CELLULAR BIOLOGY, 1991, 11 (03) :1306-1312
[10]   A CLUSTER OF PHOSPHORYLATION SITES ON THE CYCLIC AMP-REGULATED NUCLEAR FACTOR CREB PREDICTED BY ITS SEQUENCE [J].
GONZALEZ, GA ;
YAMAMOTO, KK ;
FISCHER, WH ;
KARR, D ;
MENZEL, P ;
BIGGS, W ;
VALE, WW ;
MONTMINY, MR .
NATURE, 1989, 337 (6209) :749-752