Structure and Function of Nucleoside Hydrolases from Physcomitrella patens and Maize Catalyzing the Hydrolysis of Purine, Pyrimidine, and Cytokinin Ribosides

0423120 - ÚEB 2014 RIV US eng J - Journal Article
Kopečná, M. - Blaschke, H. - Kopečný, D. - Vigouroux, A. - Končitíková, R. - Novák, Ondřej - Kotland, Ondřej - Strnad, Miroslav - Moréra, S. - von Schwartzenberg, K.
Structure and Function of Nucleoside Hydrolases from Physcomitrella patens and Maize Catalyzing the Hydrolysis of Purine, Pyrimidine, and Cytokinin Ribosides.
Plant Physiology. Roč. 163, č. 4 (2013), s. 1568-1583. ISSN 0032-0889. E-ISSN 1532-2548
Grant - others:GA MŠk(CZ) ED0007/01/01
Program: ED
Institutional research plan: CEZ:AV0Z50380511
Keywords : LUPIN LUPINUS-LUTEUS * ADENOSINE NUCLEOSIDASE * CRITHIDIA-FASCICULATA
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 7.394, year: 2013

We present a comprehensive characterization of the nucleoside N-ribohydrolase (NRH) family in two model plants, Physcomitrella patens (PpNRH) and maize (Zea mays; ZmNRH), using in vitro and in planta approaches. We identified two NRH subclasses in the plant kingdom; one preferentially targets the purine ribosides inosine and xanthosine, while the other is more active toward uridine and xanthosine. Both subclasses can hydrolyze plant hormones such as cytokinin ribosides. We also solved the crystal structures of two purine NRHs, PpNRH1 and ZmNRH3. Structural analyses, site-directed mutagenesis experiments, and phylogenetic studies were conducted to identify the residues responsible for the observed differences in substrate specificity between the NRH isoforms. The presence of a tyrosine at position 249 (PpNRH1 numbering) confers high hydrolase activity for purine ribosides, while an aspartate residue in this position confers high activity for uridine. Bud formation is delayed by knocking out single NRH genes in P. patens, and under conditions of nitrogen shortage, PpNRH1-deficient plants cannot salvage adenosine-bound nitrogen. All PpNRH knockout plants display elevated levels of certain purine and pyrimidine ribosides and cytokinins that reflect the substrate preferences of the knocked out enzymes. NRH enzymes thus have functions in cytokinin conversion and activation as well as in purine and pyrimidine metabolism.
Permanent Link: http://hdl.handle.net/11104/0229211