Global analysis reveals SRp20- and SRp75-specific mRNPs in cycling and neural cells

Minna-Liisa Ankö; Lucia Morales; Ian Henry; Andreas Beyer; Karla M Neugebauer

doi:10.1038/nsmb.1862

Global analysis reveals SRp20- and SRp75-specific mRNPs in cycling and neural cells

Minna-Liisa Ankö
, Lucia Morales
, Ian Henry
, Andreas Beyer
, Karla M Neugebauer

Biochemistry and Cell Biology

Research output: Contribution to journal › Article › Scientific › peer-review

53 Citations (Scopus)

Abstract

Members of the SR protein family of RNA-binding proteins have numerous roles in mRNA metabolism, from transcription to translation. To understand how SR proteins coordinate gene regulation, comprehensive knowledge of endogenous mRNA targets is needed. Here we establish physiological expression of GFP-tagged SR proteins from stable transgenes. Using the GFP tag for immunopurification of mRNPs, mRNA targets of SRp20 and SRp75 were identified in cycling and neurally induced P19 cells. Genome-wide analysis showed that SRp20 and SRp75 associate with hundreds of distinct, functionally related groups of transcripts that change in response to neural differentiation. Knockdown of either SRp20 or SRp75 led to up- or downregulation of specific transcripts, including identified targets, and rescue by the GFP-tagged SR proteins proved their functionality. Thus, SR proteins contribute to the execution of gene-expression programs through their association with distinct endogenous mRNAs.

Original language	English
Pages (from-to)	962-70
Number of pages	9
Journal	Nature structural & molecular biology
Volume	17
Issue number	8
DOIs	https://doi.org/10.1038/nsmb.1862
Publication status	Published - Aug 2010
MoE publication type	A1 Journal article-refereed

Keywords

Animals
Cell Cycle
Cell Differentiation/genetics
Cell Line, Tumor
Chromosomes, Artificial, Bacterial/metabolism
Gene Expression Regulation, Neoplastic
Gene Knockdown Techniques
Green Fluorescent Proteins/metabolism
Immunoprecipitation
Mice
Neurons/cytology
Organ Specificity
Protein Binding
RNA, Messenger/genetics
RNA-Binding Proteins/genetics
Ribonucleoproteins/metabolism
Serine-Arginine Splicing Factors

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10.1038/nsmb.1862

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title = "Global analysis reveals SRp20- and SRp75-specific mRNPs in cycling and neural cells",

abstract = "Members of the SR protein family of RNA-binding proteins have numerous roles in mRNA metabolism, from transcription to translation. To understand how SR proteins coordinate gene regulation, comprehensive knowledge of endogenous mRNA targets is needed. Here we establish physiological expression of GFP-tagged SR proteins from stable transgenes. Using the GFP tag for immunopurification of mRNPs, mRNA targets of SRp20 and SRp75 were identified in cycling and neurally induced P19 cells. Genome-wide analysis showed that SRp20 and SRp75 associate with hundreds of distinct, functionally related groups of transcripts that change in response to neural differentiation. Knockdown of either SRp20 or SRp75 led to up- or downregulation of specific transcripts, including identified targets, and rescue by the GFP-tagged SR proteins proved their functionality. Thus, SR proteins contribute to the execution of gene-expression programs through their association with distinct endogenous mRNAs.",

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author = "Minna-Liisa Ank{\"o} and Lucia Morales and Ian Henry and Andreas Beyer and Neugebauer, \{Karla M\}",

year = "2010",

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doi = "10.1038/nsmb.1862",

language = "English",

volume = "17",

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AU - Ankö, Minna-Liisa

AU - Morales, Lucia

AU - Henry, Ian

AU - Beyer, Andreas

AU - Neugebauer, Karla M

PY - 2010/8

Y1 - 2010/8

N2 - Members of the SR protein family of RNA-binding proteins have numerous roles in mRNA metabolism, from transcription to translation. To understand how SR proteins coordinate gene regulation, comprehensive knowledge of endogenous mRNA targets is needed. Here we establish physiological expression of GFP-tagged SR proteins from stable transgenes. Using the GFP tag for immunopurification of mRNPs, mRNA targets of SRp20 and SRp75 were identified in cycling and neurally induced P19 cells. Genome-wide analysis showed that SRp20 and SRp75 associate with hundreds of distinct, functionally related groups of transcripts that change in response to neural differentiation. Knockdown of either SRp20 or SRp75 led to up- or downregulation of specific transcripts, including identified targets, and rescue by the GFP-tagged SR proteins proved their functionality. Thus, SR proteins contribute to the execution of gene-expression programs through their association with distinct endogenous mRNAs.

AB - Members of the SR protein family of RNA-binding proteins have numerous roles in mRNA metabolism, from transcription to translation. To understand how SR proteins coordinate gene regulation, comprehensive knowledge of endogenous mRNA targets is needed. Here we establish physiological expression of GFP-tagged SR proteins from stable transgenes. Using the GFP tag for immunopurification of mRNPs, mRNA targets of SRp20 and SRp75 were identified in cycling and neurally induced P19 cells. Genome-wide analysis showed that SRp20 and SRp75 associate with hundreds of distinct, functionally related groups of transcripts that change in response to neural differentiation. Knockdown of either SRp20 or SRp75 led to up- or downregulation of specific transcripts, including identified targets, and rescue by the GFP-tagged SR proteins proved their functionality. Thus, SR proteins contribute to the execution of gene-expression programs through their association with distinct endogenous mRNAs.

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KW - Chromosomes, Artificial, Bacterial/metabolism

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KW - Green Fluorescent Proteins/metabolism

KW - Immunoprecipitation

KW - Mice

KW - Neurons/cytology

KW - Organ Specificity

KW - Protein Binding

KW - RNA, Messenger/genetics

KW - RNA-Binding Proteins/genetics

KW - Ribonucleoproteins/metabolism

KW - Serine-Arginine Splicing Factors

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DO - 10.1038/nsmb.1862

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SP - 962

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JO - Nature structural & molecular biology

JF - Nature structural & molecular biology

IS - 8

ER -

Global analysis reveals SRp20- and SRp75-specific mRNPs in cycling and neural cells

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Keywords

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