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. 1996 Oct 25;271(43):27184-7.
doi: 10.1074/jbc.271.43.27184.

Alternative splicing of human inducible nitric-oxide synthase mRNA. tissue-specific regulation and induction by cytokines

N T Eissa  1 , A J Strauss, C M Haggerty, E K Choo, S C Chu, J Moss
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Free article

Alternative splicing of human inducible nitric-oxide synthase mRNA. tissue-specific regulation and induction by cytokines

N T Eissa et al. J Biol Chem. .
Free article

Abstract

Human inducible nitric-oxide synthase (iNOS) is responsible for nitric oxide synthesis in response to inflammatory mediators. The human iNOS gene, containing 26 exons, encodes a protein of 131 kDa. This study was aimed at investigating the presence of alternative splicing of human iNOS mRNA. Total RNA from human alveolar macrophages, nasal and bronchial epithelial cells, and several human tissues was transcribed to cDNA and analyzed using polymerase chain reaction with specific primers for segmental analysis of the iNOS gene. Four sites of alternative splicing were identified by sequence analysis; these included deletion of: (i) exon 5; (ii) exons 8 and 9; (iii) exons 9, 10, and 11; and (iv) exons 15 and 16. The deduced amino acid sequences of the novel iNOS cDNAs predict one truncated protein (resulting from exon 5 deletion) and three iNOS proteins with in-frame deletions. Southern analyses of polymerase chain reaction products were consistent with tissue-specific regulation of alternative splicing. In cultured cells, iNOS induction by cytokines and lipopolysaccharide was associated with an increase in alternatively spliced mRNA transcripts. Because iNOS is active as a dimer, the novel forms of alternatively spliced iNOS may be involved in regulation of nitric oxide synthesis.

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