|, LKLF, Kruppel-like factor 2, Kruppel like factor 2|
Krüppel-like Factor 2 (KLF2), also known as lung Krüppel-like Factor (LKLF), is a protein that in humans is encoded by the KLF2 gene on chromosome 19. It is a member of the Krüppel-like factor family of zinc finger transcription factors, and it has been implicated in a variety of biochemical processes in the human body, including lung development, embryonic erythropoiesis, epithelial integrity, T-cell viability, and adipogenesis.
Erythroid Krüppel-like Factor (EKLF or KLF1) was the first Krüppel-like Factor discovered. It was found to be vitally important for embryonic erythropoiesis in promoting the switch from fetal hemoglobin (Hemoglobin F) to adult hemoglobin (Hemoglobin A) gene expression by binding to highly conserved CACCC domains. EKLF ablation in mouse embryos produces a lethal anemic phenotype, causing death by embryonic day 14, and natural mutations lead to β+ thalassemia in humans. However, expression of embryonic hemoglobin and fetal hemoglobin genes is normal in EKLF-deficient mice, and since all genes on the human β-globin locus exhibit the CACCC elements, researchers began searching for other Krüppel-like factors.
KLF2, initially called lung Krüppel-like Factor due to its high expression in the adult mouse lung, was first isolated in 1995 by using the zinc finger domain of EKLF as a hybridization probe. By transactivation assay in mouse fibroblasts, KLF2 was also noticed to bind to the β-globin gene promoter containing the CACCC sequence shown to be the binding site for EKLF, confirming KLF2 as a member of the Krüppel-like Factor family. Since then, many other KLF proteins have been discovered.
The main distinguishing feature of the KLF family is the presence of three highly conserved Cysteine2/Histidine2 zinc fingers of either 21 or 23 amino acid residues in length, located at the C-terminus of the protein. These amino acid sequences each chelate a single zinc ion, coordinated between the two cysteine and two histidine residues. These zinc fingers are joined by a conserved seven-amino acid sequence; TGEKP(Y/F)X. The zinc fingers enable all KLF proteins to bind to CACCC gene promoters, so although they may complete varied functions (due to lack of homology away from the zinc fingers), they all recognize similar binding domains.
KLF2 also exhibits these structural features. The mRNA transcript is approximately 1.5 kilobases in length, and the 37.7 kDa protein contains 354 amino acids. KLF2 also shares some homology with EKLF at the N-terminus with a proline-rich region presumed to function as the transactivation domain.
KLF2 was first discovered, and is highly expressed in, the adult mouse lung, but it is also expressed temporally during embryogenesis in erythroid cells, endothelium, lymphoid cells, the spleen, and white adipose tissue. It is expressed as early as embryonic day 9.5 in the endothelium.
KLF2 has a particularly interesting expression profile in erythroid cells. It is minimally expressed in the primitive and fetal definitive erythroid cells, but is highly expressed in adult definitive erythroid cells, particularly in the proerythroblast and the polychromatic and orthochromatic normoblasts.
Homologous recombination of embryonic stem cells was used to generate KLF2-deficient mouse embryos. Both vasculogenesis and angiogenesis were normal in the embryos, but they died by embryonic day 14.5 from severe hemorrhaging. The vasculature displayed defective morphology, with thin tunica media and aneurysmal dilation that led to rupturing. Aortic vascular smooth muscle cells failed to organize into a normal tunica media, and pericytes were low in number. These KLF2-deficient mice thus demonstrated the important role of KLF2 in blood vessel stabilization during embryogenesis.
Lung buds removed from KLF2-deficient mouse embryos and cultured from normal tracheobronchial trees. In order to circumvent embryonic lethality usually observed in KLF2-deficient embryos, KLF2 homozygous null mouse embryonic stem cells were constructed and used to produce chimeric animals. These KLF2-deficient embryonic stem cells contribute significantly to development of skeletal muscle, spleen, heart, liver, kidney, stomach, brain, uterus, testis, and skin, but not to the development of the lung. These embryos had lungs arrested in the late canalicular stage of lung development, with undilated acinar tubules. In contrast, wild type embryos are born in the saccular stage of lung development with expanded alveoli. This suggests that KLF2 is an important transcription factor required in late gestation for lung development.
KLF2 is now believed to play an important role in embryonic erythropoiesis, specifically in regulating embryonic and fetal β-like globin gene expression. In a murine KLF2-deficient embryo, expression of β-like globin genes normally expressed in primitive erythroid cells was significantly decreased, although adult β-globin gene expression was unaffected.
The role of KLF2 in human β-like globin gene expression was further elucidated by transfection of a murine KLF2-deficient embryo with the human β-globin locus. It was found that KLF2 was important for ε-globin (found in embryonic hemoglobin) and γ-globin (found in fetal hemoglobin) gene expression. However, as before, KLF2 plays no role in adult β-globin gene expression; this is regulated by EKLF.
However, KLF2 and EKLF have been found to interact in embryonic erythropoiesis. Deletion of both KLF2 and EKLF in mouse embryos results in fatal anemia earlier than in either single deletion at embryonic day 10.5. This indicates that KLF2 and EKLF interact in embryonic and fetal β-like globin gene expression. It has been shown using conditional knockout mice that both KLF2 and EKLF bind directly to β-like globin promoters. There is also evidence to suggest that KLF2 and EKLF synergistically bind to the Myc promoter, a transcription factor that is associated with gene expression of α-globin and β-globin in embryonic proerythroblasts.
This activates mechanosensitive channels, which in turn activates two pathways; the MEK5/ERK5 pathway, which activates MEF2, a transcription factor that upregulates KLF2 gene expression; and PI3K inhibition, which increases the stability of KLF2 mRNA. Binding of cytokines such as TNFα and IL-1β to their receptors activates transcription factor p65, which also induces KLF2 expression. KLF2 then has four key functions in endothelium:
- By inhibiting activation of p65 by transcription coactivator p300, VCAM1 and SELE expression is downregulated, genes that encode endothelial cell adhesion molecules, causing decreased lymphocyte and leukocyte activation and hence decreasing inflammation
- It upregulates THBD (thrombomodulin) and NOS3 (endothelial nitric oxide synthase) expression, having an anti-thrombotic effect
- Through the upregulation of NOS3, as well as NPPC (natriuretic precursor peptide C), KLF2 has a vasodilatory effect
- KLF2 also inhibits VEGFR2 (VEGF receptor 2) expression, having an anti-angiogenic effect
Thus KLF2 has an important role in regulating normal endothelium physiology. It is hypothesized that myeloid-specific KLF2 plays a protective role in atherosclerosis. Gene expression changes in endothelial cells induced by KLF2 have been demonstrated to be atheroprotective.
KLF2 has an important function in T-lymphocyte differentiation. T-cells are activated and more prone to apoptosis without KLF2, suggesting that KLF2 regulates T-cell quiescence and survival. KLF2-deficient thymocytes also do not express several receptors required for thymus emigration and differentiation into mature T-cells, such as sphingosine-1 phosphate receptor 1.
KLF2 is a negative regulator of adipocyte differentiation. KLF2 is expressed in preadipocytes, but not mature adipocytes, and it potently inhibits PPAR-γ (peroxisome proliferator-activated receptor-γ) expression by inhibiting promoter activity. This prevents differentiation of preadipocytes into adipocytes, and thus prevents adipogenesis.
- GRCh38: Ensembl release 89: ENSG00000127528 - Ensembl, May 2017
- GRCm38: Ensembl release 89: ENSMUSG00000055148 - Ensembl, May 2017
- "Human PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
- "Mouse PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
- Kozyrev SV, Hansen LL, Poltaraus AB, Domninsky DA, Kisselev LL (Jun 1999). "Structure of the human CpG-island-containing lung Krüppel-like factor (LKLF) gene and its location in chromosome 19p13.11-13 locus". FEBS Lett. 448 (1): 149–52. doi:10.1016/S0014-5793(99)00348-8. PMID 10217429.
- Wani MA, Conkright MD, Jeffries S, Hughes MJ, Lingrel JB (Sep 1999). "cDNA isolation, genomic structure, regulation, and chromosomal localization of human lung Kruppel-like factor". Genomics. 60 (1): 78–86. doi:10.1006/geno.1999.5888. PMID 10458913.
- Pearson R; Fleetwood J; Eaton S; Crossley M; Bao S (2008). "Krüppel-like transcription factors: a functional family". Int J Biochem Cell Biol. 40 (10): 1996–2001. doi:10.1016/j.biocel.2007.07.018. PMID 17904406.
- Hodge D, Coghill E, Keys J, Maguire T, Hartmann B, McDowall A, Weiss M, Grimmond S, Perkins A (April 2006). "A global role for EKLF in definitive and primitive erythropoiesis". Blood. 107 (8): 3359–70. doi:10.1182/blood-2005-07-2888. PMC 1895762. PMID 16380451.
- Perkins AC, Sharpe AH, Orkin SH (May 1995). "Lethal beta-thalassaemia in mice lacking the erythroid CACCC-transcription factor EKLF". Nature. 375 (6529): 318–22. doi:10.1038/375318a0. PMID 7753195.
- Bieker JJ (2005). "An unexpected entry into the globin real estate market". Blood. 106 (7): 2230–2231. doi:10.1182/blood-2005-07-2862.
- Anderson KP, Kern CB, Crable SC, Lingrel JB (November 1995). "Isolation of a gene encoding a functional zinc finger protein homologous to erythroid Krüppel-like factor: identification of a new multigene family". Mol. Cell. Biol. 15 (11): 5957–65. PMC 230847. PMID 7565748.
- Palis J, Kinglsey P, Stoeckert CJ. "Gene 16598: Klf2 (kruppel-like factor 2 (lung))". ErythonDB.
- Kuo CT, Veselits ML, Barton KP, Lu MM, Clendenin C, Leiden JM (November 1997). "The LKLF transcription factor is required for normal tunica media formation and blood vessel stabilization during murine embryogenesis". Genes Dev. 11 (22): 2996–3006. doi:10.1101/gad.11.22.2996. PMC 316695. PMID 9367982.
- Wani MA, Wert SE, Lingrel JB (July 1999). "Lung Kruppel-like factor, a zinc finger transcription factor, is essential for normal lung development". J. Biol. Chem. 274 (30): 21180–5. doi:10.1074/jbc.274.30.21180. PMID 10409672.
- Basu P, Morris PE, Haar JL, Wani MA, Lingrel JB, Gaensler KM, Lloyd JA (October 2005). "KLF2 is essential for primitive erythropoiesis and regulates the human and murine embryonic beta-like globin genes in vivo". Blood. 106 (7): 2566–71. doi:10.1182/blood-2005-02-0674. PMC 1895257. PMID 15947087.
- Basu P, Lung TK, Lemsaddek W, Sargent TG, Williams DC, Basu M, Redmond LC, Lingrel JB, Haar JL, Lloyd JA (November 2007). "EKLF and KLF2 have compensatory roles in embryonic β-globin gene expression and primitive erythropoiesis". Blood. 110 (9): 3417–25. doi:10.1182/blood-2006-11-057307. PMC 2200909. PMID 17675555.
- Alhashem YN, Vinjamur DS, Basu M, Klingmüller U, Gaensler KM, Lloyd JA (July 2011). "Transcription factors KLF1 and KLF2 positively regulate embryonic and fetal β-globin genes through direct promoter binding". J. Biol. Chem. 286 (28): 24819–27. doi:10.1074/jbc.M111.247536. PMC 3137057. PMID 21610079.
- Pang CJ, Lemsaddek W, Alhashem YN, Bondzi C, Redmond LC, Ah-Son N, Dumur CI, Archer KJ, Haar JL, Lloyd JA, Trudel M (July 2012). "Kruppel-like factor 1 (KLF1), KLF2, and Myc control a regulatory network essential for embryonic erythropoiesis". Mol. Cell. Biol. 32 (13): 2628–44. doi:10.1128/MCB.00104-12. PMC 3434496. PMID 22566683.
- Dekker RJ, van Soest S, Fontijn RD, Salamanca S, de Groot PG, VanBavel E, Pannekoek H, Horrevoets AJ (September 2002). "Prolonged fluid shear stress induces a distinct set of endothelial cell genes, most specifically lung Krüppel-like factor (KLF2)". Blood. 100 (5): 1689–98. doi:10.1182/blood-2002-01-0046. PMID 12176889.
- Gimbrone MA Jr, García-Cardeña G (2013). "Vascular endothelium, hemodynamics, and the pathobiology of atherosclerosis". Cardiovascular Pathology. 22 (1): 9–15. doi:10.1016/j.carpath.2012.06.006. PMC 4564111. PMID 22818581.
- Atkins GB, Jain MK (June 2007). "Role of Krüppel-like transcription factors in endothelial biology". Circ. Res. 100 (12): 1686–95. doi:10.1161/01.RES.0000267856.00713.0a. PMID 17585076.
- Shaked I, Ley K (May 2012). "Protective role for myeloid specific KLF2 in atherosclerosis". Circ. Res. 110 (10): 1266. doi:10.1161/CIRCRESAHA.112.270991. PMID 22581916.
- Carlson CM, Endrizzi BT, Wu J, Ding X, Weinreich MA, Walsh ER, Wani MA, Lingrel JB, Hogquist KA, Jameson SC (July 2006). "Kruppel-like factor 2 regulates thymocyte and T-cell migration". Nature. 442 (7100): 299–302. doi:10.1038/nature04882. PMID 16855590.
- Banerjee SS, Feinberg MW, Watanabe M, Gray S, Haspel RL, Denkinger DJ, Kawahara R, Hauner H, Jain MK (January 2003). "The Krüppel-like factor KLF2 inhibits peroxisome proliferator-activated receptor-gamma expression and adipogenesis". J. Biol. Chem. 278 (4): 2581–4. doi:10.1074/jbc.M210859200. PMID 12426306.
- KLF2+protein,+human at the US National Library of Medicine Medical Subject Headings (MeSH)
- KLF2 human gene location in the UCSC Genome Browser.
- KLF2 human gene details in the UCSC Genome Browser.