SR Protein Family Members Display Diverse Activities in the Formation of Nascent and Mature mRNPs In Vivo

Aparna K. Sapra; Minna-Liisa Änkö; Inna Grishina; Mike Lorenz; Marta Pabis; Ina Poser; Jarod Rollins; Eva Maria Weiland; Karla M. Neugebauer

doi:10.1016/j.molcel.2009.02.031

SR Protein Family Members Display Diverse Activities in the Formation of Nascent and Mature mRNPs In Vivo

Aparna K. Sapra
, Minna-Liisa Änkö
, Inna Grishina
, Mike Lorenz
, Marta Pabis
, Ina Poser
, Jarod Rollins
, Eva Maria Weiland
, Karla M. Neugebauer^*

^*Corresponding author for this work

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

122 Citations (Scopus)

Abstract

The SR proteins are a family of pre-mRNA splicing factors with additional roles in gene regulation. To investigate individual family members in vivo, we generated a comprehensive panel of stable cell lines expressing GFP-tagged SR proteins under endogenous promoter control. Recruitment of SR proteins to nascent FOS RNA was transcription dependent and RNase sensitive, with unique patterns of accumulation along the gene specified by the RNA recognition motifs (RRMs). In addition, all SR protein interactions with Pol II were RNA dependent, indicating that SR proteins are not preassembled with Pol II. SR protein interactions with RNA were confirmed in situ by FRET/FLIM. Interestingly, SC35-GFP also exhibited FRET with DNA and failed to associate with cytoplasmic mRNAs, whereas all other SR proteins underwent nucleocytoplasmic shuttling and associated with specific nuclear and cytoplasmic mRNAs. Because different constellations of SR proteins bound nascent, nuclear, and cytoplasmic mRNAs, mRNP remodeling must occur throughout an mRNA's lifetime.

Original language	English
Pages (from-to)	179-190
Number of pages	12
Journal	Molecular Cell
Volume	34
Issue number	2
DOIs	https://doi.org/10.1016/j.molcel.2009.02.031
Publication status	Published - 24 Apr 2009
Externally published	Yes
Publication type	A1 Journal article-refereed

Funding

We are grateful to I. Listerman for her early involvement in the project and comments on the manuscript. L. Morales, J. Görnemann, M. Strzelecka, and F. Carillo-Oesterreich also provided valuable insights. We thank J. Stevenin, D. Eick, E. Izuarralde, M. Carmo-Fonseca, D. Black, Y. Shav-Tal, and G. Biamonti for generous gifts of antibodies. We thank the Hyman and Buchholz labs for sharing BAC technology and M. Augsburg for help establishing the SF2- and U1-70K-tagged cell lines (with funding from Mitocheck). This work was directly supported by a Marie Curie Fellowship (to A.K.S.), a Sigrid Juselius fellowship (to M.-L.A.), the Max Planck Society, and EURASNET (FP6 Network of Excellence to K.M.N.).

Keywords

PROTEINS
RNA

ASJC Scopus subject areas

Molecular Biology
Cell Biology

Access to Document

10.1016/j.molcel.2009.02.031

Cite this

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title = "SR Protein Family Members Display Diverse Activities in the Formation of Nascent and Mature mRNPs In Vivo",

abstract = "The SR proteins are a family of pre-mRNA splicing factors with additional roles in gene regulation. To investigate individual family members in vivo, we generated a comprehensive panel of stable cell lines expressing GFP-tagged SR proteins under endogenous promoter control. Recruitment of SR proteins to nascent FOS RNA was transcription dependent and RNase sensitive, with unique patterns of accumulation along the gene specified by the RNA recognition motifs (RRMs). In addition, all SR protein interactions with Pol II were RNA dependent, indicating that SR proteins are not preassembled with Pol II. SR protein interactions with RNA were confirmed in situ by FRET/FLIM. Interestingly, SC35-GFP also exhibited FRET with DNA and failed to associate with cytoplasmic mRNAs, whereas all other SR proteins underwent nucleocytoplasmic shuttling and associated with specific nuclear and cytoplasmic mRNAs. Because different constellations of SR proteins bound nascent, nuclear, and cytoplasmic mRNAs, mRNP remodeling must occur throughout an mRNA's lifetime.",

keywords = "PROTEINS, RNA",

author = "Sapra, \{Aparna K.\} and Minna-Liisa {\"A}nk{\"o} and Inna Grishina and Mike Lorenz and Marta Pabis and Ina Poser and Jarod Rollins and Weiland, \{Eva Maria\} and Neugebauer, \{Karla M.\}",

note = "Funding Information: We are grateful to I. Listerman for her early involvement in the project and comments on the manuscript. L. Morales, J. G{\"o}rnemann, M. Strzelecka, and F. Carillo-Oesterreich also provided valuable insights. We thank J. Stevenin, D. Eick, E. Izuarralde, M. Carmo-Fonseca, D. Black, Y. Shav-Tal, and G. Biamonti for generous gifts of antibodies. We thank the Hyman and Buchholz labs for sharing BAC technology and M. Augsburg for help establishing the SF2- and U1-70K-tagged cell lines (with funding from Mitocheck). This work was directly supported by a Marie Curie Fellowship (to A.K.S.), a Sigrid Juselius fellowship (to M.-L.A.), the Max Planck Society, and EURASNET (FP6 Network of Excellence to K.M.N.). ",

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AU - Sapra, Aparna K.

AU - Änkö, Minna-Liisa

AU - Grishina, Inna

AU - Lorenz, Mike

AU - Pabis, Marta

AU - Poser, Ina

AU - Rollins, Jarod

AU - Weiland, Eva Maria

AU - Neugebauer, Karla M.

N1 - Funding Information: We are grateful to I. Listerman for her early involvement in the project and comments on the manuscript. L. Morales, J. Görnemann, M. Strzelecka, and F. Carillo-Oesterreich also provided valuable insights. We thank J. Stevenin, D. Eick, E. Izuarralde, M. Carmo-Fonseca, D. Black, Y. Shav-Tal, and G. Biamonti for generous gifts of antibodies. We thank the Hyman and Buchholz labs for sharing BAC technology and M. Augsburg for help establishing the SF2- and U1-70K-tagged cell lines (with funding from Mitocheck). This work was directly supported by a Marie Curie Fellowship (to A.K.S.), a Sigrid Juselius fellowship (to M.-L.A.), the Max Planck Society, and EURASNET (FP6 Network of Excellence to K.M.N.).

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N2 - The SR proteins are a family of pre-mRNA splicing factors with additional roles in gene regulation. To investigate individual family members in vivo, we generated a comprehensive panel of stable cell lines expressing GFP-tagged SR proteins under endogenous promoter control. Recruitment of SR proteins to nascent FOS RNA was transcription dependent and RNase sensitive, with unique patterns of accumulation along the gene specified by the RNA recognition motifs (RRMs). In addition, all SR protein interactions with Pol II were RNA dependent, indicating that SR proteins are not preassembled with Pol II. SR protein interactions with RNA were confirmed in situ by FRET/FLIM. Interestingly, SC35-GFP also exhibited FRET with DNA and failed to associate with cytoplasmic mRNAs, whereas all other SR proteins underwent nucleocytoplasmic shuttling and associated with specific nuclear and cytoplasmic mRNAs. Because different constellations of SR proteins bound nascent, nuclear, and cytoplasmic mRNAs, mRNP remodeling must occur throughout an mRNA's lifetime.

AB - The SR proteins are a family of pre-mRNA splicing factors with additional roles in gene regulation. To investigate individual family members in vivo, we generated a comprehensive panel of stable cell lines expressing GFP-tagged SR proteins under endogenous promoter control. Recruitment of SR proteins to nascent FOS RNA was transcription dependent and RNase sensitive, with unique patterns of accumulation along the gene specified by the RNA recognition motifs (RRMs). In addition, all SR protein interactions with Pol II were RNA dependent, indicating that SR proteins are not preassembled with Pol II. SR protein interactions with RNA were confirmed in situ by FRET/FLIM. Interestingly, SC35-GFP also exhibited FRET with DNA and failed to associate with cytoplasmic mRNAs, whereas all other SR proteins underwent nucleocytoplasmic shuttling and associated with specific nuclear and cytoplasmic mRNAs. Because different constellations of SR proteins bound nascent, nuclear, and cytoplasmic mRNAs, mRNP remodeling must occur throughout an mRNA's lifetime.

KW - PROTEINS

KW - RNA

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DO - 10.1016/j.molcel.2009.02.031

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AN - SCOPUS:64749099643

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JF - Molecular Cell

IS - 2

ER -

SR Protein Family Members Display Diverse Activities in the Formation of Nascent and Mature mRNPs In Vivo

Abstract

Funding

Keywords

ASJC Scopus subject areas

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