Identification of an exonic splicing silencer in exon 6A of the human VEGF gene.

BMC Molecular Biology
Rui WangNeil R Hackett

Abstract

The different isoforms of vascular endothelial growth factor (VEGF) play diverse roles in vascular growth, structure and function. Alternative splicing of the VEGF gene results in the expression of three abundant isoforms: VEGF121, VEGF165 and VEGF189. The mRNA for VEGF189 contains the alternatively spliced exon 6A whereas the mRNA for VEGF165 lacks this exon. The objective of this study was to identify the cis elements that control utilization of exon 6A. A reporter minigene was constructed (pGFP-E6A) containing the coding sequence for GFP whose translation was dependent on faithful splicing for removal of the VEGF exon 6A. To identify cis-acting splicing elements, sequential deletions were made across exon 6A in the pGFP-E6A plasmid. A candidate cis-acting exonic splicing silencer (ESS) comprising nucleotides 22-30 of exon 6A sequence was identified corresponding to the a silencer consensus sequence of AAGGGG. The function of this sequence as an ESS was confirmed in vivo both in the context of the reporter minigene as a plasmid and in the context of a longer minigene with VEGF exon 6A in its native context in an adenoviral gene transfer vector. Further mutagenesis studies resulted in the identification of the second G residue...Continue Reading

References

Nov 1, 1991·Journal of Cellular Biochemistry·N FerraraD W Leung
Jun 2, 1995·The Journal of Biological Chemistry·A P LevyM A Goldberg
Jan 1, 1993·Annual Review of Biochemistry·G DreyfussC G Burd
Mar 15, 1996·International Journal of Cancer. Journal International Du Cancer·C G KevilJ S Alexander
Oct 29, 1997·Proceedings of the National Academy of Sciences of the United States of America·S Y ChengW K Cavenee
Mar 14, 1998·Journal of the National Cancer Institute·T TokunagaY Ueyama
Aug 10, 2000·The Journal of Biological Chemistry·B L Fogel, M T McNally
Nov 30, 2000·European Journal of Cancer : Official Journal for European Organization for Research and Treatment of Cancer (EORTC) [and] European Association for Cancer Research (EACR)·Y OshikaY Ueyama
Feb 13, 2001·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Y S NgP A D'Amore
Feb 16, 2002·The EMBO Journal·Massimo Caputi, Alan M Zahler
Feb 24, 2004·The Journal of Gene Medicine·C Magin-LachmannE Wagner
Mar 11, 2004·The Journal of Biological Chemistry·Michiko MurakiMasatoshi Hagiwara
Apr 21, 2004·The Journal of Biological Chemistry·Michael HowellSharon L Milgram
Aug 6, 2004·Endocrine Reviews·Napoleone Ferrara
Nov 3, 2004·Cancer Research·Julian P Venables
Dec 21, 2004·Cell·Zefeng WangChristopher B Burge
May 21, 2005·Cellular and Molecular Life Sciences : CMLS·U Pozzoli, M Sironi
May 27, 2005·Biochemical Society Transactions·R SpellmanC W J Smith
Jul 26, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Hideki AmanoRonald G Crystal
Dec 21, 2005·Journal of Cellular and Molecular Medicine·Anne Marie ByrneJ H Harmey
Dec 31, 2005·Nucleic Acids Research·Stefan StammThangavel Alphonse Thanaraj
Oct 10, 2006·Cancer Letters·Michael R LadomeryDavid O Bates
May 2, 2007·The Journal of Gene Medicine·Olivia C HibbittRichard Wade-Martins
Jun 25, 2008·Molecular and Cellular Biology·David M MaugerMariano A Garcia-Blanco

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Citations

Sep 10, 2014·Antioxidants & Redox Signaling·Sabrina FarrokhHeiner Schaal
May 21, 2015·Einstein·Larissa Ferraz GarciaMaria Aparecida da Silva Pinhal

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Methods Mentioned

BETA
flow cytometry
transfection
transfections
transgenic
PCR
Flow Cytometric

Software Mentioned

MetaMorph

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