Sequence and expression characteristics of a nuclear-encoded chloroplast sigma factor from mustard (Sinapis alba)

被引:51
作者
Kestermann, M
Neukirchen, S
Kloppstech, K
Link, G
机构
[1] Ruhr Univ Bochum, D-44780 Bochum, Germany
[2] Leibniz Univ Hannover, Inst Bot, D-30419 Hannover, Germany
关键词
D O I
10.1093/nar/26.11.2747
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
Plant chloroplasts contain transcription factors that functionally resemble bacterial sigma factors. We have cloned the full-length cDNA from mustard (Sinapis abba) for a 53 kDa derived polypeptide that contains similarity to regions 1.2-4.2 of sigma(70)-type factors. The amino acid sequence at the N-terminus has characteristics of a chloroplast transit peptide. An in vitro synthesized polypeptide containing this region was shown to be imported into the chloroplast and processed. The recombinant factor lacking the N-terminal extension was expressed in Escherichia coli and purified. It confers the ability on E.coli core RNA polymerase to bind specifically to a DNA fragment that contains the chloroplast psbA promoter. Transcription of the psbA template by E.coli core enzyme in the presence of recombinant SIG1 results in enhanced formation of transcripts of the size expected for correct initiation at the in vivo start site. Together, these data suggest that the mature protein acts as one of the chloroplast transcription factors in mustard. RNA gel blot hybridization reveals a transcript at similar to 1.8 kb, which is more abundant in light-grown than in dark-grown mustard seedlings.
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页码:2747 / 2753
页数:7
相关论文
共 59 条
[21]  
IGLOI GL, 1992, CRIT REV PLANT SCI, V10, P525, DOI 10.1080/07352689209382326
[22]   Leaf-specifically expressed genes for polypeptides destined for chloroplasts with domains of σ70 factors of bacterial RNA polymerases in Arabidopsis thaliana [J].
Isono, K ;
Shimizu, M ;
Yoshimoto, K ;
Niwa, Y ;
Satoh, K ;
Yokota, A ;
Kobayashi, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (26) :14948-14953
[23]   SIGMA-FACTOR RELATIVES IN EUKARYOTES [J].
JAEHNING, JA .
SCIENCE, 1991, 253 (5022) :859-859
[24]  
JANG SH, 1991, J BIOL CHEM, V266, P22671
[25]  
KEEGSTRA K, 1989, ANNU REV PLANT PHYS, V40, P471, DOI 10.1146/annurev.pp.40.060189.002351
[26]  
KRUSE E, 1995, IN VITRO TRANSCRIPTI, P279
[27]   Enhanced RAGE method using specific enrichment by biotinylated oligonucleotides bound to streptavidin coated magnetic particles [J].
Lankiewicz, S ;
Gisselmann, G ;
Hatt, H .
NUCLEIC ACIDS RESEARCH, 1997, 25 (10) :2037-2038
[28]   DNA-DEPENDENT RNA-POLYMERASE OF SPINACH-CHLOROPLASTS - CHARACTERIZATION OF ALPHA-LIKE AND SIGMA-LIKE POLYPEPTIDES [J].
LERBS, S ;
BRAUTIGAM, E ;
MACHE, R .
MOLECULAR & GENERAL GENETICS, 1988, 211 (03) :459-464