INVITRO TRANSCRIPTION AND DNA-BINDING CHARACTERISTICS OF CHLOROPLAST AND ETIOPLAST EXTRACTS FROM MUSTARD (SINAPIS-ALBA) INDICATE DIFFERENTIAL USAGE OF THE PSBA PROMOTER

被引:60
作者
EISERMANN, A
TILLER, K
LINK, G
机构
关键词
DNA binding proteins; oligonucleotide-directed mutagenesis; plastid promoter; psbA gene; stage specific plastid transcription extracts;
D O I
10.1002/j.1460-2075.1990.tb07619.x
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
The psbA gene which is differentially expressed in vivo in chloroplasts and etioplasts has an unusual promoter, containing both prokaryotic-type '-35' and '-10' elements and a sequence motif that resembles the nuclear TATA box. Single base pair substitutions were introduced into the mustard psbA promoter and the mutants were tested in transcription and DNA binding experiments, using extracts from either chloroplasts or etioplasts. Positions within the '35' region appear to play an essential role in the chloroplast but not in the etioplast system. Altering the first or second position of the 'TATA box'-like region led to decreased psbA in vitro transcription in either plastid extract. These two mutations, however, did not affect binding of extracts to the (linear) psbA promoter fragment in gel retardation assays. Fragments carrying two other plastid promoters effectively competed psbA promoter binding of the etioplast extract, but more weakly that of the chloroplast extract. Lambda exonuclease mapping shows that the 5' border of the binding region is more upstream with the etioplast than with the chloroplast system, whereas the 3' border appears to be the same. Hence, protein(s) of the two plastid types seems to interact differently with the mustard psbA promoter in vitro and perhaps also in vivo.
引用
收藏
页码:3981 / 3987
页数:7
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