{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T04:08:17Z","timestamp":1790050097671,"version":"4.0.1"},"reference-count":43,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2010,1,13]],"date-time":"2010-01-13T00:00:00Z","timestamp":1263340800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Arthritis &amp; Rheumatism"],"published-print":{"date-parts":[[2010,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Objective<\/jats:title><jats:p>Scleroderma (systemic sclerosis [SSc]) is a complex connective tissue disorder characterized by hardening and thickening of the skin. One hallmark of scleroderma is excessive accumulation of collagen accompanied by increased levels of pyridinoline collagen crosslinks derived from hydroxylysine residues in the collagen telopeptide domains. Lysyl hydroxylase 2 (LH2), an important alternatively spliced enzyme in collagen biosynthesis, acts as a collagen telopeptide hydroxylase. Changes in the pattern of LH2 alternative splicing, favoring increased inclusion of the alternatively spliced LH2 exon 13A, thereby increasing the levels of the long transcript of LH2 (LH2[long]), are linked to scleroderma disease. This study was undertaken to examine the role played by RNA binding protein Fox\u20102 in regulating exon 13A inclusion, which leads to the generation of scleroderma\u2010associated LH2(long) messenger RNA (mRNA).<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Phylogenetic sequence analysis of introns flanking exon 13A was performed. A tetracycline\u2010inducible system in T\u2010Rex 293 cells was used to induce Fox\u20102 protein, and endogenous LH2(long) mRNA was determined by reverse transcriptase\u2013polymerase chain reaction. An LH2 minigene was designed, validated, and used in Fox\u20102 overexpression and mutagenesis experiments. Knockdown of Fox\u20102 was performed in mouse embryonic fibroblasts and in fibroblasts from SSc patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Overexpression of Fox\u20102 enhanced the inclusion of exon 13A and increased the generation of LH2(long) mRNA, whereas knockdown of Fox\u20102 decreased LH2(long) transcripts. Mutational analysis of an LH2 minigene demonstrated that 2 of the 4 Fox binding motifs flanking LH2 exon 13A are required for inclusion of exon 13A. In early passage fibroblasts derived from patients with scleroderma, the knockdown of Fox\u20102 protein significantly decreased the endogenous levels of LH2(long) mRNA.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Our findings indicate that Fox\u20102 plays an integral role in the regulation of LH2 splicing. Knockdown of Fox\u20102 and other methods to decrease the levels of fibrosis\u2010associated LH2(long) mRNA in primary scleroderma cells may suggest a novel approach to strategies directed against scleroderma.<\/jats:p><\/jats:sec>","DOI":"10.1002\/art.27315","type":"journal-article","created":{"date-parts":[[2010,1,13]],"date-time":"2010-01-13T15:46:09Z","timestamp":1263397569000},"page":"1167-1175","source":"Crossref","is-referenced-by-count":14,"title":["Fox\u20102 protein regulates the alternative splicing of scleroderma\u2010associated lysyl hydroxylase 2 messenger RNA"],"prefix":"10.1002","volume":"62","author":[{"given":"Puneet","family":"Seth","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heather 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