The Pitx2c N-terminal domain is a critical interaction domain required for asymmetric morphogenesis.
The paired-like homeodomain transcription factor Pitx2c has an essential role in patterning the left-right axis. However, neither its transcriptional targets nor the molecular mechanisms through which it exerts its patterning function are known. Here we provide evidence that the N-terminal domain of Pitx2c is important for this activity. Overexpression of the Pitx2c N-terminus in ovo randomizes the direction of heart looping, the first morphological asymmetry conserved in vertebrate embryos. In addition, the Pitx2c N-terminal domain blocks the ability of Pitx2c to synergize with Nkx2.5 to transactivate the procollagen lysyl hydroxylase (Plod-1) promoter in transient transfection assays. A five amino acid region containing leucine-41 is required for both of these effects. Our data suggest that the Pitx2c N-terminal domain competes with endogenous Pitx2c for binding to a protein interaction partner that is required for the activation of genes that direct asymmetric morphogenesis along the left-right axis.
Simard
Annie
A
The Research Institute of the McGill University Health Center, Montreal, Quebec, Canada.
Di Giorgio
Luciano
L
Amen
Melanie
M
Westwood
Ashley
A
Amendt
Brad A
BA
Ryan
Aimee K
AK
D000595:Amino Acid Sequence; D000818:Animals; D019521:Body Patterning / Q000502:physiology*; D002478:Cells, Cultured; D002642:Chick Embryo; D018507:Gene Expression Regulation, Developmental; D006321:Heart / Q000033:anatomy & histology / Q000196:embryology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007930:Leucine / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D000097577:Homeobox Protein PITX2
D018398:Homeodomain Proteins; D007930:Leucine; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase; D020033:Protein Isoforms; D014157:Transcription Factors; D000097577:Homeobox Protein PITX2
J Biol Chem. 2002 Jul 12;277(28):25001-10
11948188
Biochem Biophys Res Commun. 1998 Jun 9;247(1):100-5
9636662
Development. 2001 Mar;128(6):1005-13
11222154
J Cell Sci. 2005 Mar 15;118(Pt 6):1129-37
15728254
Genes Dev. 1996 May 15;10(10):1284-95
8675014
Invest Ophthalmol Vis Sci. 2006 Sep;47(9):3846-52
16936096
Mol Cell Biol. 1999 Oct;19(10):7001-10
10490637
J Cell Biol. 2001 Feb 5;152(3):545-52
11157981
Development. 1999 Mar;126(6):1225-34
10021341
Dev Biol. 2002 Jun 15;246(2):407-17
12051825
Dev Dyn. 2001 Mar;220(3):284-9
11241836
J Virol. 1987 Oct;61(10):3004-12
3041020
Mech Dev. 2005 Jan;122(1):3-25
15582774
BMC Mol Biol. 2008 Mar 28;9:31
18373856
Cell Mol Life Sci. 2000 Oct;57(11):1652-66
11092457
Development. 2001 Jun;128(11):2039-48
11493526
Development. 2000 Jul;127(14):3079-90
10862745
Circ Res. 2006 Feb 17;98(3):421-8
16397144
Cell. 1998 Aug 7;94(3):307-17
9708733
Dev Growth Differ. 2008 Jun;50 Suppl 1:S71-8
18494708
Development. 1999 Dec;126(24):5749-58
10572050
Expert Rev Mol Med. 2005 Nov 08;7(25):1-17
16274491
Hum Mol Genet. 2008 Dec 1;17(23):3643-54
18723525
Science. 1997 Feb 28;275(5304):1301-4
9036854
Nat Genet. 1996 Dec;14(4):392-9
8944018
Development. 2003 Dec;130(25):6375-85
14623826
Development. 2007 Dec;134(23):4199-208
17959721
Endocrinology. 2002 Aug;143(8):2839-51
12130547
Dev Dyn. 1992 Dec;195(4):231-72
1304821
Nature. 1998 Aug 6;394(6693):545-51
9707115
Development. 2002 Jan;129(2):329-37
11807026
Proc Natl Acad Sci U S A. 2008 Aug 12;105(32):11242-7
18678914
Cell. 1998 Aug 7;94(3):319-24
9708734
J Biol Chem. 1998 Aug 7;273(32):20066-72
9685346
J Biol Chem. 2003 Jun 20;278(25):22437-45
12692125
Cell. 1998 Aug 7;94(3):299-305
9708732
Birth Defects Res A Clin Mol Teratol. 2006 Mar;76(3):175-81
16498627
Development. 2008 Feb;135(4):677-85
18199582
Development. 1999 Oct;126(20):4643-51
10498698
Hum Mol Genet. 1997 Mar;6(3):457-64
9147650
Mech Dev. 2000 Jan;90(1):41-51
10585561
Development. 2000 Mar;127(5):1081-93
10662647
J Neurochem. 2000 Nov;75(5):1818-25
11032870
Hum Mol Genet. 2002 May 15;11(10):1177-84
12015277
Dev Biol. 2006 Aug 15;296(2):437-49
16836994
Mech Dev. 2006 Mar;123(3):210-27
16500087
Neuroendocrinology. 2000 May;71(5):277-86
10859489
J Dent Res. 2003 Dec;82(12):1008-12
14630904
Mol Cell Biol. 2007 Nov;27(21):7560-73
17785445
Nature. 1999 Sep 16;401(6750):279-82
10499586
Dev Cell. 2008 Jul;15(1):134-45
18606147
J Biol Chem. 2004 Dec 10;279(50):52087-94
15466416
Development. 2002 Nov;129(21):5081-91
12397115
Mol Vis. 2008;14:2205-10
19052653
Nature. 1999 Sep 16;401(6750):276-8
10499585
Dev Biol. 2003 Oct 15;262(2):268-81
14550790
J Biol Chem. 2001 Jun 22;276(25):23034-41
11301317