HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 35, 36660-36669, August 27, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/35/36660    most recent M405380200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ellis, P. D. Articles by Kemp, P. Search for Related Content PubMed PubMed Citation Articles by Ellis, P. D. Articles by Kemp, P. Social Bookmarking                      What's this?

Regulated Tissue-specific Alternative Splicing of Enhanced Green Fluorescent Protein Transgenes Conferred by -Tropomyosin Regulatory Elements in Transgenic Mice^*

Peter D. Ellis, Christopher W. J. Smith, and Paul Kemp

From the Department of Biochemistry, Tennis Court Road, University of Cambridge, Cambridge CB2 1QW, United Kingdom

The mutually exclusive exons 2 and 3 of -tropomyosin (TM) have been used as a model system for strictly regulated alternative splicing. Exon 2 inclusion is only observed at high levels in smooth muscle (SM) tissues, whereas striated muscle and non-muscle cells use predominantly exon 3. Experiments in cell culture have shown that exon 2 selection results from repression of exon 3 and that this repression is mediated by regulatory elements flanking exon 3. We have now tested the cell culture-derived model in transgenic mice. We show that by harnessing the intronic splicing regulatory elements, expression of an enhanced green fluorescent protein transgene with a constitutively active promoter can be restricted to SM cells. Splicing of both endogenous TM and a series of transgenes carrying regulatory element mutations was analyzed by reverse transcriptasePCR. These studies indicated that although SM-rich tissues are equipped to regulate splicing of high levels of endogenous or transgene TM RNA, other non-SM tissues such as spleen, which express lower amounts of TM, also splice significant proportions of exon 2, and this splicing pattern can be recapitulated by transgenes expressed at low levels. We confirm the importance in vivo of the negatively acting regulatory elements for regulated skipping of exon 3. Moreover, we provide evidence that some of the regulatory factors responsible for exon 3 skipping appear to be titratable, with loss of regulated splicing sometimes being associated with high transgene expression levels.

Received for publication, May 14, 2004

^* This work was supported by Project Grant PG/2001031 from the British Heart Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence may be addressed. Tel.: 44-1223-333655; Fax: 44-1223-766002; E-mail: cwjs1{at}cam.ac.uk. To whom correspondence may be addressed. Tel.: 44-1223-333627; Fax: 44-1223-766002; E-mail: pk{at}mole.bio.cam.ac.uk.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				A. R. Gordon, S. V. Outram, M. Keramatipour, C. A. Goddard, W. H. Colledge, J. C. Metcalfe, A. L. Hager-Theodorides, T. Crompton, and P. R. Kemp Splenomegaly and Modified Erythropoiesis in KLF13-/- Mice J. Biol. Chem., May 2, 2008; 283(18): 11897 - 11904. [Abstract] [Full Text] [PDF]

				P. Gunning, G. O'neill, and E. Hardeman Tropomyosin-Based Regulation of the Actin Cytoskeleton in Time and Space Physiol Rev, January 1, 2008; 88(1): 1 - 35. [Abstract] [Full Text] [PDF]

				T. Moroy and F. Heyd The impact of alternative splicing in vivo: Mouse models show the way RNA, August 1, 2007; 13(8): 1155 - 1171. [Abstract] [Full Text] [PDF]

				J. P. Orengo, D. Bundman, and T. A. Cooper A bichromatic fluorescent reporter for cell-based screens of alternative splicing Nucleic Acids Res., December 2, 2006; 34(22): e148 - e148. [Abstract] [Full Text] [PDF]

				J. B. Crawford and J. G. Patton Activation of {alpha}-Tropomyosin Exon 2 Is Regulated by the SR Protein 9G8 and Heterogeneous Nuclear Ribonucleoproteins H and F Mol. Cell. Biol., December 1, 2006; 26(23): 8791 - 8802. [Abstract] [Full Text] [PDF]

				V. I. Bonano, S. Oltean, R. M. Brazas, and M. A. Garcia-Blanco Imaging the alternative silencing of FGFR2 exon IIIb in vivo RNA, December 1, 2006; 12(12): 2073 - 2079. [Abstract] [Full Text] [PDF]

				E. A. Newman, S. J. Muh, R. H. Hovhannisyan, C. C. Warzecha, R. B. Jones, W. L. McKeehan, and R. P. Carstens Identification of RNA-binding proteins that regulate FGFR2 splicing through the use of sensitive and specific dual color fluorescence minigene assays RNA, June 1, 2006; 12(6): 1129 - 1141. [Abstract] [Full Text] [PDF]

				S.-N. Grellscheid and C. W. J. Smith An Apparent Pseudo-Exon Acts both as an Alternative Exon That Leads to Nonsense-Mediated Decay and as a Zero-Length Exon. Mol. Cell. Biol., March 1, 2006; 26(6): 2237 - 2246. [Abstract] [Full Text] [PDF]

				L. Favot, S. M. Hall, S. G. Haworth, and P. R. Kemp Cytoplasmic YY1 Is Associated with Increased Smooth Muscle-Specific Gene Expression: Implications for Neonatal Pulmonary Hypertension Am. J. Pathol., December 1, 2005; 167(6): 1497 - 1509. [Abstract] [Full Text] [PDF]