Integrin linked kinase (ILK) is a scaffold protein, which plays important roles in hair follicle development. The cDNA sequence of novel ILK gene in sheep was cloned by PCR method and analyzed by bioinformatics. Tissue expression profiling in eight tissues and temporal profiling at different wool follicle anagen stages in skin was analyzed. The results showed that the whole open reading frame (ORF) of ILK gene was 1 359 bp in length, which encoded 452 amino acids. Bioinformatic analysis indicated that the secondary structure of ILK gene was mainly made up of three ankyrin repeats and a kinase domain, and there were multiple phosphorylation and Protein Kinase C sites in this gene. The RT-PCR result confirmed that ILK mRNA was expressed in heart, liver, spleen, lung, skeletal muscle, skin, and small intestine, and the expression level was much higher in skin, spleen, and liver than others. The q-PCR analysis demon-strated that the expression level of ILK was significantly increased from March to May (early follicle anagen initia-tion) in both sheep breeds, Chinese Merino and Kazakh sheep, and there were certain differences from June to October be-tween the two breeds. The above results indicated that ILK gene may play key roles in regulating secondary follicle growth.
YANG Jian-Bo, GAN Shang-Quan, YANG Yong-Lin, ZHANG Hong-Lin, SONG Tian-Zeng, FENG Jing, YANG Jing-Quan, GAO Lei, SHI Guo-Qing, SHEN Min
. Cloning and expression in follicle anagen of ILK gene in sheep[J]. Hereditas(Beijing), 2012
, 34(6)
: 719
-726
.
DOI: 10.3724/SP.J.1005.2012.00719
[1] Hannigan GE, Leung-Hagesteijn C, Fitz-Gibbon L, Coppolino MG, Radeva G, Filmus J, Bell JC, Dedhar S. Regulation of cell adhesion and anchorage-dependent growth by a new β1-integrin-linked protein kinase. Nature, 1996, 379(6560): 91-96.
[2] Delcommenne M, Tan C, Gray V, Rue L, Woodgett J, Dedhar S. Phosphoinositide-3-OH kinase-dependent regulation of glycogen synthase kinase 3 and protein kinase B/AKT by the integrin-linked kinase. Proc Natl Acad Sci USA, 1998, 95(19): 11211-11216.
[3] Novak A, Hsu SC, Leung-Hagesteijn C, Radeva G, Papkoff J, Montesano R, Roskelley C, Grosschedl R, Dedhar S. Cell adhesion and the integrin-linked kinase regulate the LEF-1 and β-catenin signaling pathways. Proc Natl Acad Sci USA, 1998, 95(8): 4374-4379.
[4] Haase M, Gmach CC, Eke I, Hehlgans S, Baretton GB, Cordes N. Expression of integrin-linked kinase is increased in differentiated cells. J Histochem Cytochem, 2008, 56(9): 819-829.
[5] Takanami I. Increased expression of integrin-linked kinase is associated with shorter survival in non-small cell lung cancer. BMC Cancer, 2005, 5: 1.
[6] Nakrieko KA, Welch I, Dupuis H, Bryce D, Pajak A, St Arnaud R, Dedhar S, D'Souza SJ, Dagnino L. Impaired hair follicle morphogenesis and polarized keratinocyte movement upon conditional inactivation of integrin-linked kinase in the epidermis. Mol Biol Cell, 2008, 19(4): 1462-1473.
[7] Xie W, Li FG, Kudlow JE, Wu CY. Expression of the integrin-linked kinase (ILK) in mouse skin. Am J Pathol, 1998, 153(2): 367-372.
[8] Lorenz K, Grashoff C, Torka R, Sakai T, Langbein L, Bloch W, Aumailley M, Fassler R. Integrin-linked kinase is required for epidermal and hair follicle morphogenesis. J Cell Biol, 2007, 177(3): 501-513.
[9] Wickstrom SA, Lange A, Hess MW, Polleux J, Spatz JP, Kruger M, Pfaller K, Lambacher A, Bloch W, Mann M, Huber LA, Fassler R. Integrin-linked kinase controls microtubule dynamics required for plasma membrane targeting of caveolae. Dev Cell, 2010, 19(4): 574-588.
[10] Vi L, de Lasa C, DiGuglielmo GM, Dagnino L. Integrin- linked kinase is required for TGF-β1 induction of dermal myofibroblast differentiation. J Invest Dermatol, 2011, 131(3): 586-593.
[11] 赵伯阳, 汪代华, 徐刚毅, 赵文伯, 郑程莉. 山羊CAST基因Ⅱ型转录本的克隆及在山羊不同组织中的表达. 遗传, 2011, 33(4): 358-364.
[12] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 2001, 25(4): 402-408.
[13] Legate KR, Montanez E, Kudlacek O, Fassler R. ILK, PINCH and parvin: the tIPP of integrin signalling. Nat Rev Mol Cell Biol, 2006, 7(1): 20-31.
[14] Grashoff C. The role of Integrin-linked kinase in vivo and in virto
[Degree thesis]. The Ludwig-Maximilian University, 2007.
[15] Hannigan G, Troussard AA, Dedhar S. Integrin-linked kinase: a cancer therapeutic target unique among its ILK. Nat Rev Cancer, 2005, 5(1): 51-63.
[16] Chiswell BP, Zhang R, Murphy JW, Boggon TJ, Calder-wood DA. The structural basis of integrin-linked kinase-PINCH interactions. Proc Natl Acad Sci USA, 2008, 105(52): 20677-20682.
[17] Lange A, Wickstrom SA, Jakobson M, Zent R, Sainio K, Fassler R. Integrin-linked kinase is an adaptor with essential functions during mouse development. Nature, 2009, 461 (7266): 1002-1006.
[18] 赵有璋. 羊生产学 (第二版). 北京: 中国农业出版社, 2003: 32-36.
[19] Nakrieko KA, Rudkouskaya A, Irvine TS, D'Souza SJ, Dagnino L. Targeted inactivation of integrin-linked kinase in hair follicle stem cells reveals an important modulatory role in skin repair after injury. Mol Biol Cell, 2011, 22(14): 2532-2540.
