Alternate processing of Flt1 transcripts is directed by conserved cis-elements within an intronic region of FLT1 that reciprocally regulates splicing and polyadenylation

Christie P Thomas; Nandita S Raikwar; Elizabeth A Kelley; Kang Z Liu

doi:10.1093/nar/gkq198

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Alternate processing of Flt1 transcripts is directed by conserved cis-elements within an intronic region of FLT1 that reciprocally regulates splicing and polyadenylation

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Alternate processing of Flt1 transcripts is directed by conserved cis-elements within an intronic region of FLT1 that reciprocally regulates splicing and polyadenylation

Christie P Thomas, Nandita S Raikwar, Elizabeth A Kelley and Kang Z Liu

Nucleic acids research, Vol.38(15), pp.5130-5140

08/2010

DOI: 10.1093/nar/gkq198

PMCID: PMC2926593

PMID: 20385595

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Abstract

The vascular endothelial growth factor receptor, Flt1 is a transmembrane receptor co-expressed with an alternate transcript encoding a secreted form, sFlt1, that functions as a competitive inhibitor of Flt1. Despite shared transcription start sites and upstream regulatory elements, sFlt1 is in far greater excess of Flt1 in the human placenta. Phorbol myristic acid and dimethyloxalylglycine differentially stimulate sFlt1 compared to Flt1 expression in vascular endothelial cells and in cytotrophoblasts. An FLT1 minigene construct containing exon 13, 14 and the intervening region, recapitulates mRNA processing when transfected into COS-7, with chimeric intronic sFlt1 transcripts arising by intronic polyadenylation and other Flt1/sFlt1 transcripts by alternate splicing. Inclusion of exon 15 but not 14 had a modest stimulatory effect on the abundance of sFlt1. The intronic region containing the distal poly(A) signal sequences, when transferred to a heterologous minigene construct, inhibited splicing but only when cloned in sense orientation, consistent with the presence of a directional cis-element. Serial deletional and targeted mutational analysis of cis-elements within intron 13 identified intronic poly(A) signal sequences and adjacent cis-elements as the principal determinants of the relative ratio of intronic sFlt1 and spliced Flt1. We conclude that intronic signals reciprocally regulate splicing and polyadenylation and control sFlt1 expression.

Endothelium, Vascular - cytology

Trophoblasts - metabolism

Alternative Splicing

Introns

Humans

Cells, Cultured

Cercopithecus aethiops

Regulatory Sequences, Ribonucleic Acid

Vascular Endothelial Growth Factor Receptor-1 - metabolism

Animals

Endothelium, Vascular - metabolism

Base Sequence

Polyadenylation

Conserved Sequence

COS Cells

Vascular Endothelial Growth Factor Receptor-1 - genetics

Details

Title: Subtitle: Alternate processing of Flt1 transcripts is directed by conserved cis-elements within an intronic region of FLT1 that reciprocally regulates splicing and polyadenylation
Creators: Christie P Thomas - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA, USA. christie-thomas@uiowa.edu
Nandita S Raikwar
Elizabeth A Kelley
Kang Z Liu
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.38(15), pp.5130-5140
DOI: 10.1093/nar/gkq198
PMID: 20385595
PMCID: PMC2926593
NLM abbreviation: Nucleic Acids Res
ISSN: 0305-1048
eISSN: 1362-4962
Publisher: England
Grant note: HL71664 / NHLBI NIH HHS DK52617 / NIDDK NIH HHS R01 DK090053 / NIDDK NIH HHS
Language: English
Date published: 08/2010
Academic Unit: Stead Family Department of Pediatrics; Obstetrics and Gynecology; Internal Medicine
Record Identifier: 9983986082002771

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