Intracellular localization of Arabidopsis sulfurtransferases

被引:35
作者
Bauer, M
Dietrich, C
Nowak, K
Sierralta, WD
Papenbrock, J [1 ]
机构
[1] Leibniz Univ Hannover, Inst Bot, D-30419 Hannover, Germany
[2] Leibniz Univ Hannover, Inst Plant Dis & Plant Protect, D-30419 Hannover, Germany
[3] Tech Univ Braunschweig, Inst Plant Biol, D-38106 Braunschweig, Germany
[4] Univ Chile, Lab Ultrastruct, INTA, Santiago, Chile
关键词
D O I
10.1104/pp.104.040121
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Sulfurtransferases (Str) comprise a group of enzymes widely distributed in archaea, eubacteria, and eukaryota which catalyze the transfer of a sulfur atom from suitable sulfur donors to nucleophilic sulfur acceptors. In all organisms analyzed to date, small gene families encoding Str proteins have been identified. The gene products were localized to different compartments of the cells. Our interest concerns the localization of Str proteins encoded in the nuclear genome of Arabidopsis. Computer-based prediction methods revealed localization in different compartments of the cell for six putative AtStrs. Several methods were used to determine the localization of the AtStr proteins experimentally. For AtStr1, a mitochondrial localization was demonstrated by immunodetection in the proteome of isolated mitochondria resolved by one- and two-dimensional gel electrophoresis and subsequent blotting. The respective mature AtStr1 protein was identified by mass spectrometry sequencing. The same result was obtained by transient expression of fusion constructs with the green fluorescent protein in Arabidopsis protoplasts, whereas AtStr2 was exclusively localized to the cytoplasm by this method. Three members of the single-domain AtStr were localized in the chloroplasts as demonstrated by transient expression of green fluorescent protein fusions in protoplasts and stomata, whereas the single-domain AtStr18 was shown to be cytoplasmic. The remarkable subcellular distribution of AtStr15 was additionally analyzed by transmission electron immunomicroscopy using a monospecific antibody against green fluorescent protein, indicating an attachment to the thylakoid membrane. The knowledge of the intracellular localization of the members of this multiprotein family will help elucidate their specific functions in the organism.
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页码:916 / 926
页数:11
相关论文
共 41 条
[1]  
Abel S, 1998, METH MOL B, V82, P209
[2]   Identification and characterization of single-domain thiosulfate sulfurtransferases from Arabidopsis thaliana [J].
Bauer, M ;
Papenbrock, J .
FEBS LETTERS, 2002, 532 (03) :427-431
[3]   RHODANESE-MEDIATED SULFUR TRANSFER TO SUCCINATE-DEHYDROGENASE [J].
BONOMI, F ;
PAGANI, S ;
CERLETTI, P ;
CANNELLA, C .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1977, 72 (01) :17-24
[4]   The rhodanese/Cdc25 phosphatase superfamily - Sequence-structure-function relations [J].
Bordo, D ;
Bork, P .
EMBO REPORTS, 2002, 3 (08) :741-746
[5]   SEEKING A BETTER JOB FOR AN UNDER-EMPLOYED ENZYME - RHODANESE [J].
CERLETTI, P .
TRENDS IN BIOCHEMICAL SCIENCES, 1986, 11 (09) :369-372
[6]   EFFICIENT TRANSFORMATION OF ARABIDOPSIS-THALIANA USING DIRECT GENE-TRANSFER TO PROTOPLASTS [J].
DAMM, B ;
SCHMIDT, R ;
WILLMITZER, L .
MOLECULAR AND GENERAL GENETICS, 1989, 217 (01) :6-12
[7]   DISRUPTION OF A RHODANESELIKE GENE RESULTS IN CYSTEINE AUXOTROPHY IN SACCHAROPOLYSPORA-ERYTHRAEA [J].
DONADIO, S ;
SHAFIEE, A ;
HUTCHINSON, CR .
JOURNAL OF BACTERIOLOGY, 1990, 172 (01) :350-360
[8]   Predicting subcellular localization of proteins based on their N-terminal amino acid sequence [J].
Emanuelsson, O ;
Nielsen, H ;
Brunak, S ;
von Heijne, G .
JOURNAL OF MOLECULAR BIOLOGY, 2000, 300 (04) :1005-1016
[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]   Proteomic analysis of leaf peroxisomal proteins in greening cotyledons of Arabidopsis thaliana [J].
Fukao, Y ;
Hayashi, M ;
Nishimura, M .
PLANT AND CELL PHYSIOLOGY, 2002, 43 (07) :689-696