Document Detail


Identification of 2'-phosphodiesterase, which plays a role in the 2-5A system regulated by interferon.
MedLine Citation:
PMID:  15231837     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The 2-5A system is one of the major pathways for antiviral and antitumor functions that can be induced by interferons (IFNs). The 2-5A system is modulated by 5'-triphosphorylated, 2',5'-phosphodiester-linked oligoadenylates (2-5A), which are synthesized by 2',5'-oligoadenylate synthetases (2',5'-OASs), inactivated by 5'-phosphatase and completely degraded by 2'-phosphodiesterase (2'-PDE). Generated 2-5A activates 2-5A-dependent endoribonuclease, RNase L, which induces RNA degradation in cells and finally apoptosis. Although 2',5'-OASs and RNase L have been molecularly cloned and studied well, the identification of 2'-PDE has remained elusive. Here, we describe the first identification of 2'-PDE, the third key enzyme of the 2-5A system. We found a putative 2'-PDE band on SDS-PAGE by successive six-step chromatographies from ammonium sulfate precipitates of bovine liver and identified a partial amino acid sequence of the human 2'-PDE by mass spectrometry. Based on the full-length sequence of the human 2'-PDE obtained by in silico expressed sequence tag assembly, the gene was cloned by reverse transcription-PCR. The recombinant human 2'-PDE expressed in mammalian cells certainly cleaved the 2',5'-phosphodiester bond of 2-5A trimer and 2-5A analogs. Because no sequences with high homology to this human 2'-PDE were found, the human 2'-PDE was considered to be a unique enzyme without isoform. Suppression of 2'-PDE by a small interfering RNA and a 2'-PDE inhibitor resulted in significant reduction of viral replication, whereas overexpression of 2'-PDE protected cells from IFN-induced antiproliferative activity. These observations identify 2'-PDE as a key regulator of the 2-5A system and as a potential novel target for antiviral and antitumor treatments.
Authors:
Kazuishi Kubota; Kaori Nakahara; Toshiaki Ohtsuka; Shuku Yoshida; Junko Kawaguchi; Yoko Fujita; Yohei Ozeki; Ayako Hara; Chigusa Yoshimura; Hidehiko Furukawa; Hideyuki Haruyama; Kimihisa Ichikawa; Makoto Yamashita; Tatsuji Matsuoka; Yasuteru Iijima
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Publication Detail:
Type:  Journal Article     Date:  2004-06-30
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  279     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2004 Sep 
Date Detail:
Created Date:  2004-08-30     Completed Date:  2004-12-10     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  37832-41     Citation Subset:  IM    
Affiliation:
Biomedical Research Laboratories, Sankyo Co., Ltd., 1-2-58, Hiromachi, Shinagawa-ku, Tokyo 140-8710, Japan. kkubot@sankyo.co.jp
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AB115695
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MeSH Terms
Descriptor/Qualifier:
Adenine Nucleotides / metabolism*
Amino Acid Sequence
Animals
Base Sequence
Cattle
DNA Primers
Exoribonucleases / antagonists & inhibitors,  chemistry,  metabolism*
Hela Cells
Humans
Liver / enzymology
Molecular Sequence Data
Oligoribonucleotides / metabolism*
Phosphodiesterase Inhibitors / pharmacology
Sequence Homology, Amino Acid
Virus Replication / drug effects
Chemical
Reg. No./Substance:
0/Adenine Nucleotides; 0/DNA Primers; 0/Oligoribonucleotides; 0/Phosphodiesterase Inhibitors; 61172-40-5/2',5'-oligoadenylate; EC 3.1.-/Exoribonucleases; EC 3.1.13.-/2'-phosphodiesterase

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