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Molecular and Cellular Biology, October 2008, p. 6033-6043, Vol. 28, No. 19
0270-7306/08/$08.00+0     doi:10.1128/MCB.00726-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Multiple and Specific mRNA Processing Targets for the Major Human hnRNP Proteins{triangledown} ,{dagger}

Julian P. Venables,1 Chu-Shin Koh,1 Ulrike Froehlich,1 Elvy Lapointe,1 Sonia Couture,1 Lyna Inkel,1 Anne Bramard,1 Éric R. Paquet,1 Valérie Watier,1 Mathieu Durand,1 Jean-François Lucier,1 Julien Gervais-Bird,1 Karine Tremblay,1 Panagiotis Prinos,1 Roscoe Klinck,1,2 Sherif Abou Elela,1,2 and Benoit Chabot1,2*

Laboratoire de Génomique Fonctionnelle de l'Université de Sherbrooke,1 Département de Microbiologie et d'Infectiologie, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, Sherbrooke, Québec J1H 5N4, Canada2

Received 6 May 2008/ Returned for modification 30 May 2008/ Accepted 11 July 2008

Alternative splicing is a key mechanism regulating gene expression, and it is often used to produce antagonistic activities particularly in apoptotic genes. Heterogeneous nuclear ribonucleoparticle (hnRNP) proteins form a family of RNA-binding proteins that coat nascent pre-mRNAs. Many but not all major hnRNP proteins have been shown to participate in splicing control. The range and specificity of hnRNP protein action remain poorly documented, even for those affecting splice site selection. We used RNA interference and a reverse transcription-PCR screening platform to examine the implications of 14 of the major hnRNP proteins in the splicing of 56 alternative splicing events in apoptotic genes. Out of this total of 784 alternative splicing reactions tested in three human cell lines, 31 responded similarly to a knockdown in at least two different cell lines. On the other hand, the impact of other hnRNP knockdowns was cell line specific. The broadest effects were obtained with hnRNP K and C, two proteins whose role in alternative splicing had not previously been firmly established. Different hnRNP proteins affected distinct sets of targets with little overlap even between closely related hnRNP proteins. Overall, our study highlights the potential contribution of all of these major hnRNP proteins in alternative splicing control and shows that the targets for individual hnRNP proteins can vary in different cellular contexts.

* Corresponding author. Mailing address: Département de Microbiologie et d'Infectiologie, Faculté de Médecine et des Sciences de la Santé, 3001, 12th Avenue Nord, Université de Sherbrooke, Sherbrooke, Québec J1H 5N4, Canada. Phone: (819) 564-5321. Fax: (819) 564-5392. E-mail: Benoit.Chabot{at}USherbrooke.ca

{triangledown} Published ahead of print on 21 July 2008.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, October 2008, p. 6033-6043, Vol. 28, No. 19
0270-7306/08/$08.00+0     doi:10.1128/MCB.00726-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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