The AtNFS2 gene from Arabidopsis thaliana encodes a NifS-like plastildial cysteine desulphurase

被引:83
作者
Léon, S
Touraine, B
Briat, JF
Lobréaux, S
机构
[1] Univ Montpellier 2, Inst Natl Rech Agron, CNRS, Unite Mixte Rech 5004, F-34060 Montpellier 1, France
[2] Ecole Natl Super Agron Montpellier, F-34060 Montpellier 1, France
关键词
desulfurase; iron-sulphur cluster; plastid; thale cress; thiamin;
D O I
10.1042/BJ20020322
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
NifS-like proteins are cysteine desulphurases required for the mobilization of sulphur from cysteine. They are present in all organisms, where they are involved in iron-sulphur (Fe-S) cluster biosynthesis. In eukaryotes, these enzymes are present in mitochondria, which are the major site for Fe-S cluster assembly. The genome of the model plant Arabidopsis thaliana contains two putative NifS-like proteins. A cDNA corresponding to one of them was cloned by reverse-transcription PCR, and named AtNFS2. The corresponding transcript is expressed in many plant tissues. It encodes a protein highly related (75 % similarity) to the slr0077-gene product from Synechocystis PCC 6803, and is predicted to be targeted to plastids. Indeed, a chimaeric AtNFS2-GFP fusion protein, containing one-third of AtNFS2 from its N-terminal end, was addressed to chloroplasts. Overproduction in Escherichia coli and purification of recombinant AtNFS2 protein enabled one to demonstrate that it bears a pyridoxal 5'-phosphate-dependent cysteine desulphurase activity in vitro, thus being the first NifS homologue characterized to date in plants. The putative physiological functions of this gene are discussed, including the attractive hypothesis of a possible role in Fe-S cluster assembly in plastids.
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页码:557 / 564
页数:8
相关论文
共 49 条
[1]  
ALTSCHUL SF, 1990, J MOL BIOL, V215, P403, DOI 10.1006/jmbi.1990.9999
[2]   Comparison of the sequences of the Aspergillus nidulans hxB and Drosophila melanogaster ma-I genes with nifS from Azotobacter vinelandii suggests a mechanism for the insertion of the terminal sulphur atom in the molybdopterin cofactor [J].
Amrani, L ;
Primus, J ;
Glatigny, A ;
Arcangeli, L ;
Scazzocchio, C ;
Finnerty, V .
MOLECULAR MICROBIOLOGY, 2000, 38 (01) :114-125
[3]  
AXELOS M, 1992, PLANT PHYSIOL BIOCH, V30, P123
[4]   Evidence for the thiamine biosynthetic pathway in higher-plant plastids and its developmental regulation [J].
Belanger, FC ;
Leustek, T ;
Chu, BY ;
Kriz, AL .
PLANT MOLECULAR BIOLOGY, 1995, 29 (04) :809-821
[5]   ABA3 is a molybdenum cofactor sulfurase required for activation of aldehyde oxidase and xanthine dehydrogenase in Arabidopsis thaliana [J].
Bittner, F ;
Oreb, M ;
Mendel, RR .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (44) :40381-40384
[6]   Dual targeting properties of the N-terminal signal sequence of Arabidopsis thaliana THI1 protein to mitochondria and chloroplasts [J].
Chabregas, SM ;
Luche, DD ;
Farias, LP ;
Ribeiro, AF ;
van Sluys, MA ;
Menck, CFM ;
Silva-Filho, MC .
PLANT MOLECULAR BIOLOGY, 2001, 46 (06) :639-650
[7]   Soluble, highly fluorescent variants of green fluorescent protein (GFP) for use in higher plants [J].
Davis, SJ ;
Vierstra, RD .
PLANT MOLECULAR BIOLOGY, 1998, 36 (04) :521-528
[8]   ChloroP, a neural network-based method for predicting chloroplast transit peptides and their cleavage sites [J].
Emanuelsson, O ;
Nielsen, H ;
Von Heijne, G .
PROTEIN SCIENCE, 1999, 8 (05) :978-984
[9]   Prediction of organellar targeting signals [J].
Emanuelsson, O ;
von Heijne, G .
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, 2001, 1541 (1-2) :114-119
[10]   A modular domain of NifU, a nitrogen fixation cluster protein, is highly conserved in evolution [J].
Hwang, DM ;
Dempsey, A ;
Tan, KT ;
Liew, CC .
JOURNAL OF MOLECULAR EVOLUTION, 1996, 43 (05) :536-540