Molecular cloning, polymorphism, and expression analysis of the LKB1/STK11 gene and its association with non-specific digestive disorder in rabbits
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Liver kinase B1 (LKB1, also called STK11) encodes a serine/threonine kinase mutated in Peutz–Jeghers cancer syndrome characterized by gastrointestinal polyposis. Although LKB1 plays an important role in regulating energy homeostasis, cell growth, and metabolism via activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK), nothing is known about its molecular characteristics and possible involvement in non-specific digestive disorder (NSDD) of rabbits. In the present study, we first cloned the coding sequence (CDS) of rabbit LKB1, which consisted of 1317 bp encoding 438 amino acids (AAs) and contained a highly conserved S_TKc kinase domain. Its deduced AA sequence showed 87.93–91.10% similarities with that of other species. In order to determine its involvement in NSDD, a NSDD rabbit model was built by a dietary fiber deficiency. The polymorphic site of LKB1 was then investigated in both healthy and NSDD groups using directing sequencing. Our results suggested that a synonymous variant site (840 c. G > C, CCC→CCG) existed in its S_TKc domain, which was associated with susceptibility to NSDD. Furthermore, qPCR was utilized to examine the mRNA levels of LKB1 and its downstream targets (i.e., PRKAA2, mTOR and NF-kβ) in several intestinal-related tissues from both healthy and NSDD groups. Significant changes in their expression levels between two groups indicated that impaired LKB1 signaling contributed to the intestinal abnormality in NSDD rabbits. Taken together, it could be concluded that LKB1 might be a potential candidate gene affecting the occurrence of rabbit NSDD. This information may serve as a basis for further investigations on rabbit digestive diseases.
KeywordsLKB1 Molecular cloning Polymorphism Non-specific digestive disorder Rabbit
This study was financially supported by the 12th Five-Year Breeding Research projects in Sichuan (Grant No. 2011NZ0099-4), China Agricultural Research System (Grant No. CARS-44-A-2), and the double-support project of Sichuan Agricultural University.
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Conflict of interest
The authors have declared that no conflict of interest exists.
- 2.Trocino A, Fragkiadakis M, Majolini D, Tazzoli M, Radaelli G, Xiccato G (2013) Soluble fibre, starch and protein level in diets for growing rabbits: effects on digestive efficiency and productive traits. Anim Feed Sci Tech 180(1–4):73–82. https://doi.org/10.1016/j.anifeedsci.2013.01.007 CrossRefGoogle Scholar
- 5.Su GH, Hruban RH, Bansal RK, Bova GS, Tang DJ, Shekher MC, Westerman AM, Entius MM, Goggins M, Yeo CJ (1999) Germline and somatic mutations of the STK11/LKB1 Peutz-Jeghers gene in pancreatic and biliary cancers. Am J Pathol 154:1835–1840. https://doi.org/10.1016/S0002-9440(10)65440-5 CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Hawley SA, Boudeau J, Reid JL, Mustard KJ, Udd L, Mäkelä TP, Alessi DR, Hardie D (2003) Complexes between the LKB1 tumor suppressor, STRADα/β and MO25α/β are upstream kinases in the AMP-activated protein kinase cascade. J Biol 2(4):28. https://doi.org/10.1186/1475-4924-2-28 CrossRefPubMedPubMedCentralGoogle Scholar
- 25.Ho PA, Kuhn J, Gerbing RB, Pollard JA, Zeng R, Miller KL, Heerema NA, Raimondi SC, Hirsch BA, Franklin JL (2011) WT1 synonymous single nucleotide polymorphism rs16754 correlates with higher mRNA expression and predicts significantly improved outcome in favorable-risk pediatric acute myeloid leukemia: a report from the Children’s Oncology Group. J Clin Oncol 29:704–711. https://doi.org/10.1200/JCO.2010.31.9327 CrossRefPubMedGoogle Scholar
- 26.Stark K, Esslinger UB, Reinhard W, Petrov G, Winkler T, Komajda M, Isnard R, Charron P, Villard E, Cambien F (2010) Genetic association study identifies HSPB7 as a risk gene for idiopathic dilated cardiomyopathy. PLoS Genet 6:e1001167. https://doi.org/10.1371/journal.pgen.1001167 CrossRefPubMedPubMedCentralGoogle Scholar
- 29.Shaw RJ, Kosmatka M, Bardeesy N, Hurley RL, Witters LA, Depinho RA, Cantley LC (2004) The tumor suppressor LKB1 kinase directly activates AMP-activated kinase and regulates apoptosis in response to energy stress. Proc Natl Acad Sci USA 101:3329–3335. https://doi.org/10.1073/pnas.0308061100 CrossRefPubMedPubMedCentralGoogle Scholar