Exon organization and novel alternative splicing of the human ANK2 gene: implications for cardiac function and human cardiac disease.
Recent findings illustrate a critical role for ankyrin-B function in normal cardiovascular physiology. Specifically, decreased expression of ankyrin-B in mice or human mutations in the ankyrin-B gene (ANK2) results in potentially fatal cardiac arrhythmias. Despite the clear role of ankyrin-B in heart, the mechanisms underlying transcriptional regulation of ANK2 are unknown. In fact, to date there is no description of ANK2 genomic organization. The aims of this study were to provide a comprehensive description of the ANK2 gene and to evaluate the relative expression of alternative splicing events associated with ANK2 transcription in heart. Using reverse-transcriptase PCR on mRNA isolated from human hearts, we identify seven new exons associated with the ANK2 gene including an alternative first exon located approximately 145 kb upstream of the previously-identified first exon. In addition, we identify over thirty alternative splicing events associated with ANK2 mRNA transcripts. Using real-time PCR and exon boundary-spanning primers to selectively amplify these splice variants, we demonstrate that these variants are expressed at varying levels in human heart. Finally, ankyrin-B immunoblot analysis demonstrates the expression of a heterogeneous population of ankyrin-B polypeptides in heart. ANK2 consists of 53 exons that span approximately 560 kb on human chromosome 4. Additionally, our data demonstrates that ANK2 is subject to complex transcriptional regulation that likely results in differential ankyrin-B polypeptide function.
Cunha
Shane R
SR
Department of Internal Medicine, Division of Cardiovascular Medicine, University of Iowa Carver College of Medicine, Iowa City, IA 52242, USA. shane-cunha@uiowa.edu
Le Scouarnec
Solena
S
Schott
Jean-Jacques
JJ
Mohler
Peter J
PJ
D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D017487:Ankyrins / Q000096:biosynthesis* / Q000235:genetics; D001145:Arrhythmias, Cardiac / Q000235:genetics / Q000378:metabolism*; D002894:Chromosomes, Human, Pair 4 / Q000235:genetics / Q000378:metabolism; D005091:Exons / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D009124:Muscle Proteins / Q000096:biosynthesis* / Q000235:genetics; D009206:Myocardium / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics
C498994:ANK2 protein, human; D017487:Ankyrins; D009124:Muscle Proteins; D012333:RNA, Messenger
Nature. 1990 Jun 21;345(6277):736-9
2141669
Circulation. 2007 Jan 30;115(4):432-41
17242276
J Biol Chem. 1987 Aug 5;262(22):10537-45
3038887
J Cell Biol. 1996 May;133(4):819-30
8666667
Nucleic Acids Res. 2001 Jul 1;29(13):E68-8
11433044
J Biol Chem. 2006 Mar 3;281(9):5741-9
16368689
J Cell Sci. 1997 May;110 ( Pt 10):1239-49
9191047
N Engl J Med. 1990 Oct 11;323(15):1046-50
1977081
J Cell Biol. 1998 Nov 30;143(5):1295-304
9832557
J Biol Chem. 2007 Nov 2;282(44):32384-96
17720975
J Biol Chem. 1999 Dec 10;274(50):35908-13
10585476
Mol Cell Neurosci. 2002 May;20(1):43-55
12056839
J Biol Chem. 1995 Dec 29;270(52):31298-302
8537399
Cell. 2004 Oct 15;119(2):257-72
15479642
J Cell Biol. 1997 Mar 10;136(5):1059-70
9060470
Cardiovasc Res. 1997 Jul;35(1):2-3
9302340
Exp Cell Res. 2005 Sep 10;309(1):86-98
15953600
J Biol Chem. 2004 Jun 11;279(24):25798-804
15075330
Nat Genet. 1996 Jun;13(2):214-8
8640229
Am J Physiol. 1998 Jan;274(1):F129-38
9458832
Proc Natl Acad Sci U S A. 2004 Dec 14;101(50):17533-8
15579534
J Biol Chem. 1998 Jan 16;273(3):1339-48
9430667
J Biol Chem. 2007 Feb 16;282(7):4875-4883
17178715
J Cell Biol. 1991 Jul;114(2):241-53
1830053
J Biol Chem. 2004 Sep 17;279(38):40185-93
15262991
J Biol Chem. 1993 May 5;268(13):9533-40
8486643
J Biol Chem. 1998 Nov 13;273(46):30785-94
9804856
Physiol Rev. 2001 Jul;81(3):1353-92
11427698
J Cell Biol. 1995 Dec;131(6 Pt 2):1821-9
8557748
Proc Natl Acad Sci U S A. 1990 Apr;87(8):3117-21
2139228
J Cell Biol. 2001 Nov 26;155(5):739-46
11724816
Proc Natl Acad Sci U S A. 2004 Jun 15;101(24):9137-42
15178757
Nucleic Acids Res. 2001 May 1;29(9):e45
11328886
Exp Cell Res. 2006 Nov 1;312(18):3546-58
16962094
J Biol Chem. 1992 Sep 15;267(26):18966-72
1388161
J Biol Chem. 2002 Apr 12;277(15):12978-87
11796721
PLoS Biol. 2005 Dec;3(12):e423
16292983
J Cell Biol. 1998 Nov 30;143(5):1305-15
9832558
J Cell Biol. 1997 Feb 10;136(3):621-31
9024692
J Biol Chem. 1990 Jun 25;265(18):10589-96
2141335
J Cell Sci. 2007 Mar 15;120(Pt 6):953-63
17311847
J Biol Chem. 2004 Mar 26;279(13):12980-7
14722080
J Cell Biol. 1999 Nov 29;147(5):995-1008
10579720
Nature. 2003 Feb 6;421(6923):634-9
12571597
J Cell Biochem. 2008 Jul 1;104(4):1244-53
18275062
Am J Physiol Heart Circ Physiol. 2007 Jul;293(1):H109-19
17416611
J Biol Chem. 2002 Mar 22;277(12):10599-607
11781319
Genomics. 1998 May 15;50(1):79-88
9628825
J Cell Biol. 1998 Sep 21;142(6):1571-81
9744885
Cardiovasc Res. 2006 Jul 1;71(1):22-9
16650839