THE GENERALLY EXPRESSED HNRNP-F IS INVOLVED IN A NEURAL-SPECIFIC PRE-MESSENGER-RNA SPLICING EVENT

被引:174
作者
MIN, HS
CHAN, RC
BLACK, DL
机构
[1] UNIV CALIF LOS ANGELES, INST MOLEC BIOL, LOS ANGELES, CA 90024 USA
[2] UNIV CALIF LOS ANGELES, DEPT MICROBIOL & MOLEC GENET, LOS ANGELES, CA 90024 USA
[3] UNIV CALIF LOS ANGELES, HOWARD HUGHES MED INST, LOS ANGELES, CA 90024 USA
关键词
SPLICING; HNRNP F; NEURAL-SPECIFIC SPLICING; RNA REGULATORY COMPLEXES;
D O I
10.1101/gad.9.21.2659
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The proteins and RNA regulatory elements that control tissue-specific pre-mRNA splicing in mammalian cells are mostly unknown. In this study, a set of proteins is identified that binds to a splicing regulatory element downstream of the neuron specific c-src N1 exon. This complex of proteins bound specifically to a short RNA containing the regulatory sequence in neuronal extracts that splice the N1 exon. It was not seen in non-neuronal cell extracts that fail to splice this exon. W-cross-linking experiments identified a neuron-specific 75-kD protein and several nontissue-specific proteins, including the 53-kD heterogeneous nuclear ribonucleoprotein F (hnRNP F), as components of this complex. Although present in both extracts, hnRNP F binds tightly to the RNA only in the neuronal extracts. A mutation in the regulatory RNA sequence, that inhibits N1 splicing in vivo, abolished formation of the neuron-specific complex and the binding of the neuron-specific 75-kD protein. Competition experiments in the two extracts show that the binding of the neuronal protein complex to the src pre-mRNA is required to activate N1 exon splicing in vitro. Antibody inhibition experiments indicate that the hnRNP F protein is a functional part of this complex. The assembly of regulatory complexes from both constitutive and specific proteins is likely to be a general feature of tissue-specific splicing regulation.
引用
收藏
页码:2659 / 2671
页数:13
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