Alternative promoter activation leads to the expression of a novel human lysyl oxidase variant that functions as an amine oxidase

Kim,Seonkwan; Park,Sunhyang; Kim,Youngho

doi:10.3892/ijmm.2014.1845

Alternative promoter activation leads to the expression of a novel human lysyl oxidase variant that functions as an amine oxidase

Authors:
- Seonkwan Kim
- Sunhyang Park
- Youngho Kim
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Affiliations: Department of Biochemistry, Wonkwang University School of Medicine, Iksan, Jeollabuk-Do 570-749, Republic of Korea
Published online on: July 10, 2014 https://doi.org/10.3892/ijmm.2014.1845
Pages: 894-899

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Abstract

The lysyl oxidase (LOX) family is an emerging family of amine oxidases that is responsible for lysine-mediated crosslinks found in collagen and elastin. Several novel functions, such as tumor suppression, tumor progression, cellular senescence, chemotaxis and the modification of histones have recently been attributed to the LOX family of proteins. In the search for more human LOX paralogs, in the present study, we identified several expressed sequence tag (EST) clones that showed an alternative exon-intron splice pattern from LOX. These ESTs corresponded to the LOX transcript variant 2 (LOX-v2) that was recently reported in GenBank (accession no. NM_001178102). LOX-v2 mRNA lacks exon 1 of LOX, but contains an additional 222 bp sequence from the 5'-flanking intronic region of exon 2. The deduced LOX-v2 polypeptide contains the characteristic C-terminal domains of the LOX family, but does not contain the N-terminal propeptide region that has been reported to have tumor suppressor activity. In peroxidase-coupled fluorometric assays, LOX-v2 showed β-aminopropionitrile-inhibitable amine oxidase activity toward collagen and elastin. RT-PCR analysis of human tissues revealed a distinct tissue specificity of LOX-v2 expression compared to that of LOX. Promoter assays indicated that an alternative promoter element present in the exon 1 region of LOX was sufficient for the differential expression of LOX-v2. These findings indicate that the human LOX gene encodes 2 variants, LOX and LOX-v2, both of which function as amine oxidases with distinct tissue specificities.

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1	Kagan HM and Trackman PC: Properties and function of lysyl oxidase. Am J Respir Cell Mol Biol. 5:206–210. 1991. View Article : Google Scholar : PubMed/NCBI
2	Smith-Mungo LI and Kagan HM: Lysyl oxidase: Properties, regulation and multiple functions in biology. Matrix Biol. 16:387–398. 1998. View Article : Google Scholar : PubMed/NCBI
3	Hämäläinen ER, Jones TA, Sheer D, Taskinen K, Pihlajaniemi T and Kivirikko KI: Molecular cloning of human lysyl oxidase and assignment of the gene to chromosome 5q23.3–31.2. Genomics. 11:508–516. 1991.PubMed/NCBI
4	Kim Y, Boyd CD and Csiszar K: A new gene with sequence and structural similarity to the gene encoding human lysyl oxidase. J Biol Chem. 270:7176–7182. 1995. View Article : Google Scholar : PubMed/NCBI
5	Jourdan-Le Saux C, Tronecker H, Bogic L, Bryant-Greenwood GD, Boyd CD and Csiszar K: The LOXL2 gene encodes a new lysyl oxidase-like protein and is expressed at high levels in reproductive tissues. J Biol Chem. 274:12939–12944. 1999.PubMed/NCBI
6	Mäki JM and Kivirikko KI: Cloning and characterization of a fourth human lysyl oxidase isoenzyme. Biochem J. 355:381–387. 2001.PubMed/NCBI
7	Asuncion L, Fogelgren B, Fong KS, Fong SF, Kim Y and Csiszar K: A novel human lysyl oxidase-like gene (LOXL4) on chromosome 10q24 has an altered scavenger receptor cysteine rich domain. Matrix Biol. 20:487–491. 2001. View Article : Google Scholar : PubMed/NCBI
8	Kenyon K, Contente S, Trackman PC, Tang J, Kagan HM and Friedman RM: Lysyl oxidase and rrg messenger RNA. Science. 253:8021991. View Article : Google Scholar : PubMed/NCBI
9	Giampuzzi M, Botti G, Cilli M, Gusmano R, Borel A, Sommer P and Di Donato A: Down-regulation of lysyl oxidase-induced tumorigenic transformation in NRK-49F cells characterized by constitutive activation of ras proto-oncogene. J Biol Chem. 276:29226–29232. 2001. View Article : Google Scholar
10	Erler JT, Bennewith KL, Nicolau M, Dornhöfer N, Kong C, Le QT, Chi JT, Jeffrey SS and Giaccia AJ: Lysyl oxidase is essential for hypoxia-induced metastasis. Nature. 440:1222–1226. 2006. View Article : Google Scholar : PubMed/NCBI
11	Schietke R, Warnecke C, Wacker I, Schödel J, Mole DR, Campean V, Amann K, Goppelt-Struebe M, Behrens J, Eckardt KU and Wiesener MS: The lysyl oxidases LOX and LOXL2 are necessary and sufficient to repress E-cadherin in hypoxia: insights into cellular transformation processes mediated by HIF-1. J Biol Chem. 285:6658–6669. 2010. View Article : Google Scholar : PubMed/NCBI
12	Peinado H, Del Carmen Iglesias-de la Cruz M, Olmeda D, Csiszar K, Fong KS, Vega S, Nieto MA, Cano A and Portillo F: A molecular role for lysyl oxidase-like 2 enzyme in snail regulation and tumor progression. EMBO J. 24:3446–3458. 2005. View Article : Google Scholar : PubMed/NCBI
13	Lucero HA, Ravid K, Grimsby JL, Rich CB, DiCamillo SJ, Mäki JM, Myllyharju J and Kagan HM: Lysyl oxidase oxidizes cell membrane proteins and enhances the chemotactic response of vascular smooth muscle cells. J Biol Chem. 283:24103–24117. 2008. View Article : Google Scholar : PubMed/NCBI
14	Kagan HM, Williams MA, Calaman SD and Berkowitz EM: Histone H1 is a substrate for lysyl oxidase and contains endogenous sodium borotritide-reducible residues. Biochem Biophys Res Commun. 115:186–192. 1983. View Article : Google Scholar : PubMed/NCBI
15	Giampuzzi M, Oleggini R and Di Donato A: Demonstration of in vitro interaction between tumor suppressor lysyl oxidase and histones H1 and H2: definition of the regions involved. Biochim Biophys Acta. 1647:245–251. 2003. View Article : Google Scholar : PubMed/NCBI
16	Herranz N, Dave N, Millanes-Romero A, Morey L, Díaz VM, Lórenz-Fonfría V, Gutierrez-Gallego R, Jerónimo C, Di Croce L, García de Herreros A and Peiró S: Lysyl oxidase-like 2 deaminates lysine 4 in histone H3. Mol Cell. 46:369–376. 2012. View Article : Google Scholar : PubMed/NCBI
17	Jung ST, Kim MS, Seo JY, Kim HC and Kim Y: Purification of enzymatically active human lysyl oxidase and lysyl oxidase-like protein from Escherichia coli inclusion bodies. Protein Expr Purif. 31:240–246. 2003. View Article : Google Scholar : PubMed/NCBI
18	Palamakumbura AH and Trackman PC: A fluorometric assay for detection of lysyl oxidase enzyme activity in biological samples. Anal Biochem. 300:245–251. 2002. View Article : Google Scholar : PubMed/NCBI
19	Lee JE and Kim Y: A tissue-specific variant of the human lysyl oxidase-like protein 3 (LOXL3) functions as an amine oxidase with substrate specificity. J Biol Chem. 281:37282–37290. 2006. View Article : Google Scholar : PubMed/NCBI
20	Kim MS, Kim SS, Jung ST, Park JY, Yoo HW, Ko J, Csiszar K, Choi SY and Kim Y: Expression and purification of enzymatically active forms of the human lysyl oxidase-like protein 4. J Biol Chem. 278:52071–52074. 2003. View Article : Google Scholar : PubMed/NCBI
21	Pinnell SR and Martin G: The cross-linking of collagen and elastin: enzymatic conversion of lysine in peptide linkage to alpha-aminoadipic-delta-semialdehyde (allysine) by an extract from bone. Proc Natl Acad Sci USA. 61:708–716. 1968. View Article : Google Scholar : PubMed/NCBI
22	Kagan HM, Sullivan KA, Olsson TA III and Cronlund AL: Purification and properties of four species of lysyl oxidase from bovine aorta. Biochem J. 177:203–214. 1979.PubMed/NCBI
23	Sullivan KA and Kagan HM: Evidence for structural similarities in the multiple forms of aortic and cartilage lysyl oxidase and a catalytically quiescent aortic protein. J Biol Chem. 257:13520–13526. 1982.PubMed/NCBI
24	Kuivaniemi H, Savolainen ER and Kivirikko KI: Human placental lysyl oxidase. Purification, partial characterization, and preparation of two specific antisera to the enzyme. J Biol Chem. 259:6996–7002. 1984.PubMed/NCBI
25	Li W, Nellaiappan K, Strassmaier T, Graham L, Thomas KM and Kagan HM: Localization and activity of lysyl oxidase within nuclei of fibrogenic cells. Proc Natl Acad Sci USA. 94:12817–12822. 1997. View Article : Google Scholar : PubMed/NCBI
26	Cronshaw AD, Fothergill-Gilmore LA and Hulmes DJS: The proteolytic processing site of the precursor of lysyl oxidase. Biochem J. 306:279–284. 1995.PubMed/NCBI
27	Hayashi K, Fong KS, Mercier F, Boyd CD, Csiszar K and Hayashi M: Comparative immunocytochemical localization of lysyl oxidase (LOX) and the lysyl oxidase-like (LOXL) proteins: changes in the expression of LOXL during development and growth of mouse tissues. J Mol Histol. 35:845–855. 2004. View Article : Google Scholar
28	Mello ML, Contente S, Vidal BC, Planding W and Schenck U: Modulation of ras transformation affecting chromatin supraorganization as assessed by image analysis. Exp Cell Res. 220:374–382. 1995. View Article : Google Scholar : PubMed/NCBI
29	Kirschmann DA, Seftor EA, Nieva DR, Mariano EA and Hendrix MJ: Differentially expressed genes associated with the metastatic phenotype in breast cancer. Breast Cancer Res Treat. 55:127–136. 1999. View Article : Google Scholar : PubMed/NCBI
30	Stassar MJ, Devitt G, Brosius M, Rinnab L, Prang J, Schradin T, Simon J, Petersen S, Kopp-Schneider A and Zoller M: Identification of human renal cell carcinoma associated genes by suppression subtractive hybridization. Br J Cancer. 85:1372–1382. 2001. View Article : Google Scholar : PubMed/NCBI
31	Ren C, Yang G, Timme TL, Wheeler TM and Thompson TC: Reduced lysyl oxidase messenger RNA levels in experimental and human prostate cancer. Cancer Res. 58:1285–1290. 1998.PubMed/NCBI
32	Woznick AR, Braddock AL, Dulai M, Seymour ML, Callahan RE, Welsh RJ, Chmielewski GW, Zelenock GB and Shanley CJ: Lysyl oxidase expression in bronchogenic carcinoma. Am J Surg. 189:297–301. 2005. View Article : Google Scholar : PubMed/NCBI
33	Kaneda A, Wakazono K, Tsukamoto T, Watanabe N, Yagi Y, Tatematsu M, Kaminishi M, Sugimura T and Ushijima T: Lysyl oxidase is a tumor suppressor gene inactivated by methylation and loss of heterozygosity in human gastric cancers. Cancer Res. 64:6410–6415. 2004. View Article : Google Scholar : PubMed/NCBI
34	Kirschmann DA, Seftor EA, Fong SF, Nieva DR, Sullivan CM, Edwards EM, Sommer P, Csiszar K and Hendrix MJ: A molecular role for lysyl oxidase in breast cancer invasion. Cancer Res. 62:4478–4483. 2002.PubMed/NCBI
35	Palamakumbura AH, Jeay S, Guo Y, Pischon N, Sommer P, Sonenshein GE and Trackman PC: The propeptide domain of lysyl oxidase induces phenotypic reversion of ras-transformed cells. J Biol Chem. 279:40593–40600. 2004. View Article : Google Scholar : PubMed/NCBI
36	Min C, Yu Z, Kirsch KH, Zhao Y, Vora SR, Trackman PC, Spicer DB, Rosenberg L, Palmer JR and Sonenshein GE: A loss-of-function polymorphism in the propeptide domain of the LOX gene and breast cancer. Cancer Res. 69:6685–6693. 2009. View Article : Google Scholar : PubMed/NCBI
37	Zhao Y, Min C, Vora SR, Trackman PC, Sonenshein GE and Kirsch KH: The lysyl oxidase pro-peptide attenuates fibronectin-mediated activation of focal adhesion kinase and p130Cas in breast cancer cells. J Biol Chem. 284:1385–1393. 2009. View Article : Google Scholar : PubMed/NCBI
38	Palamakumbura AH, Vora SR, Nugent MA, Kirsch KH, Sonenshein GE and Trackman PC: Lysyl oxidase propeptide inhibits prostate cancer cell growth by mechanisms that target FGF-2-cell binding and signaling. Oncogene. 28:3390–3400. 2009. View Article : Google Scholar : PubMed/NCBI