Determination of the RNA binding specificity of the heterogeneous nuclear ribonucleoprotein (hnRNP) H/H′/F/2H9 family

被引:168
作者
Caputi, M
Zahler, AM [1 ]
机构
[1] Univ Calif Santa Cruz, Dept Mol & Cellular & Dev Biol, Santa Cruz, CA 95064 USA
[2] Univ Calif Santa Cruz, Ctr Mol Biol RNA, Sinsheimer Labs, Santa Cruz, CA 95064 USA
关键词
D O I
10.1074/jbc.M102861200
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
Members of the heterogeneous nuclear ribonucleoprotein (hnRNP) H protein family, H, H', F, and 2H9, are involved in pre-mRNA processing. We analyzed the assembly of these proteins from splicing extracts onto four RNA regulatory elements as follows: a high affinity hnRNP Al-binding site (WA1), a sequence involved in Rev-dependent export (p17gag INS), an exonic splicing silencer from the beta -tropomyosin gene, and an intronic splicing regulator (downstream control sequence (DCS) from the c-src gene. The entire family binds the WA1, instability (INS), and beta -tropomyosin substrates, and the core-binding site for each is a run of three G residues followed by an A. Transfer of small regions containing this sequence to a substrate lacking hnRNP H binding activity is sufficient to promote binding of all family members. The c-src DCS has been shown to assemble hnRNP H, not hnRNP F, from HeLa cell extracts, and we show that hnRNP 2H9 does not bind this element. The DCS contains five G residues followed by a C. Mutation of the C to an A changes the specificity of the DCS from a substrate that binds only hnRNP H/H' to a binding site for all hnRNP H family members. We conclude that the sequence GGGA is recognized by all hnRNP H family proteins.
引用
收藏
页码:43850 / 43859
页数:10
相关论文
共 58 条
[1]  
Ashiya M, 1997, RNA, V3, P996
[2]   DSEF-1 is a member of the hnRNP H family of RNA-binding proteins and stimulates pre-mRNA cleavage and polyadenylation in vitro [J].
Bagga, PS ;
Arhin, GK ;
Wilusz, J .
NUCLEIC ACIDS RESEARCH, 1998, 26 (23) :5343-5350
[3]   ACTIVATION OF C-SRC NEURON-SPECIFIC SPLICING BY AN UNUSUAL RNA ELEMENT INVIVO AND INVITRO [J].
BLACK, DL .
CELL, 1992, 69 (05) :795-807
[4]   CONSERVED STRUCTURES AND DIVERSITY OF FUNCTIONS OF RNA-BINDING PROTEINS [J].
BURD, CG ;
DREYFUSS, G .
SCIENCE, 1994, 265 (5172) :615-621
[5]   RNA-BINDING SPECIFICITY OF HNRNP A1 - SIGNIFICANCE OF HNRNP A1 HIGH-AFFINITY BINDING-SITES IN PRE-MESSENGER-RNA SPLICING [J].
BURD, CG ;
DREYFUSS, G .
EMBO JOURNAL, 1994, 13 (05) :1197-1204
[6]   hnRNP A/B proteins are required for inhibition of HIV-1 pre-mRNA splicing [J].
Caputi, M ;
Mayeda, A ;
Krainer, AR ;
Zahler, AM .
EMBO JOURNAL, 1999, 18 (14) :4060-4067
[7]   hnRNP A1 selectively interacts through its Gly-rich domain with different RNA-binding proteins [J].
Cartegni, L ;
Maconi, M ;
Morandi, E ;
Cobianchi, F ;
Riva, S ;
Biamonti, G .
JOURNAL OF MOLECULAR BIOLOGY, 1996, 259 (03) :337-348
[8]   An intron element modulating 5' splice site selection in the hnRNP A1 pre-mRNA interacts with hnRNP A1 [J].
Chabot, B ;
Blanchette, M ;
Lapierre, I ;
LaBranche, H .
MOLECULAR AND CELLULAR BIOLOGY, 1997, 17 (04) :1776-1786
[9]  
CHAN RC, 1995, MOL CELL BIOL, V15, P6377
[10]   The polypyrimidine tract binding protein binds upstream of neural cell-specific c-src exon N1 to repress the splicing of the intron downstream [J].
Chan, RCC ;
Black, DL .
MOLECULAR AND CELLULAR BIOLOGY, 1997, 17 (08) :4667-4676