Sulfate reduction in higher plants: Molecular evidence for a novel 5'-adenylylsulfate reductase

被引:131
作者
Setya, A [1 ]
Murillo, M [1 ]
Leustek, T [1 ]
机构
[1] RUTGERS STATE UNIV,DEPT PLANT SCI,CTR AGR MOL BIOL,NEW BRUNSWICK,NJ 08903
关键词
D O I
10.1073/pnas.93.23.13383
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Sulfate-assimilating organisms reduce inorganic sulfate for Cys biosynthesis, There are two leading hypotheses for the mechanism of sulfate reduction in higher plants. In one, adenosine 5'-phosphosulfate (APS) (5'-adenylylsulfate) sulfotransferase carries out reductive transfer of sulfate from APS to reduced glutathione. Alternatively, the mechanism may be similar to that in bacteria in which the enzyme, 3'-phosphoadenosine-5'-phosphosulfate (PAPS) reductase, catalyzes thioredoxin (Trx)-dependent reduction of PAPS. Three classes of cDNA were cloned from Arabidopsis thaliana termed APR1, -2, and -3, that functionally complement a cysH, PAPS reductase mutant strain of Escherichia coli. The coding sequence of the APR clones is homologous with PAPS reductases from microorganisms. In addition, a carboxyl-terminal domain is homologous with members of the Trx superfamily. Further genetic analysis showed that the APR clones can functionally complement a mutant strain of E. coli lacking Trx, and an APS kinase, cysC. mutant, These results suggest that the APR enzyme may be a Trx-independent APS reductase, Cell extracts of E. coli expressing APR showed Trx-independent sulfonucleotide reductase activity with a preference for APS over PAPS as a substrate, APR-mediated APS reduction is dependent on dithiothreitol, has a pH optimum of 8.5, is stimulated by high ionic strength, and is sensitive to inactivation by 5'-adenosinemonophosphate (5'-AMP). 2'-AMP, or 3'-phosphoadenosine-5'-phosphate (PAP), a competitive inhibitor of PAPS reductase, do not affect activity, The APR enzymes may be localized in different cellular compartments as evidenced by the presence of an amino-terminal transit peptide for plastid localization in APR1 and APR3 but not APR2. Southern blot analysis confirmed that the APR clones are members of a small gene family, possibly consisting of three members.
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页码:13383 / 13388
页数:6
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