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פותח על ידי קלירמאש פתרונות בע"מ -
Chlamydomonas reinhardtii encodes a single sigma70-like factor which likely functions in chloroplast transcription
Year:
2006
Source of publication :
Current Genetics
Authors :
יריחימוביץ', ורד
;
.
Volume :
49
Co-Authors:
Bohne, A.-V., Institut für Biologie, Humboldt-Universität, Chausseestr. 117, 10115 Berlin, Germany, Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14853, United States
Irihimovitch, V., Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14853, United States
Weihe, A., Institut für Biologie, Humboldt-Universität, Chausseestr. 117, 10115 Berlin, Germany
Stern, D.B., Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14853, United States
Facilitators :
From page:
333
To page:
340
(
Total pages:
8
)
Abstract:
Chlamydomonas reinhardtii EST clones encoding a protein highly similar to prokaryotic sigma factors and plant sigma-like factors (SLFs) were used to isolate a BAC clone containing the full-length gene CrRpoD. The gene is likely to be single-copy, in contrast to small gene families encoding SLFs in plants. The CrRpoD mRNA comprises 3,033 nt with an open reading frame of 2,256 nt, encoding a putative protein of 752 amino acids with a molecular mass of 80.2 kDa. The sequence contains conserved regions 2-4 typically found in sigma factors, and an unusually long amino terminal extension, which by in silico analysis has properties of a chloroplast transit peptide. Expression of CrRpoD was confirmed by immunodetection of a 85 kDa polypeptide in a preparation enriched for chloroplast proteins. To demonstrate functionality in transcription initiation, a recombinant CrRpoD-thioredoxin fusion protein was reconstituted with E. coli RNA polymerase core enzyme and tested in vitro. This chimeric holoenzyme specifically bound the spinach psbA and Chlamydomonas rrn16 promoters in gel mobility shift assays and exhibited specific transcription initiation from the same two promoters, providing evidence for the role of CrRpoD as a functional transcription factor. © Springer-Verlag 2006.
Note:
Related Files :
Animals
Chlamydomonas
computer model
DNA-Directed RNA Polymerases
multigene family
Spinacia oleracea
עוד תגיות
תוכן קשור
More details
DOI :
10.1007/s00294-006-0060-7
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
18737
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:23
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Scientific Publication
Chlamydomonas reinhardtii encodes a single sigma70-like factor which likely functions in chloroplast transcription
49
Bohne, A.-V., Institut für Biologie, Humboldt-Universität, Chausseestr. 117, 10115 Berlin, Germany, Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14853, United States
Irihimovitch, V., Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14853, United States
Weihe, A., Institut für Biologie, Humboldt-Universität, Chausseestr. 117, 10115 Berlin, Germany
Stern, D.B., Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14853, United States
Chlamydomonas reinhardtii encodes a single sigma70-like factor which likely functions in chloroplast transcription
Chlamydomonas reinhardtii EST clones encoding a protein highly similar to prokaryotic sigma factors and plant sigma-like factors (SLFs) were used to isolate a BAC clone containing the full-length gene CrRpoD. The gene is likely to be single-copy, in contrast to small gene families encoding SLFs in plants. The CrRpoD mRNA comprises 3,033 nt with an open reading frame of 2,256 nt, encoding a putative protein of 752 amino acids with a molecular mass of 80.2 kDa. The sequence contains conserved regions 2-4 typically found in sigma factors, and an unusually long amino terminal extension, which by in silico analysis has properties of a chloroplast transit peptide. Expression of CrRpoD was confirmed by immunodetection of a 85 kDa polypeptide in a preparation enriched for chloroplast proteins. To demonstrate functionality in transcription initiation, a recombinant CrRpoD-thioredoxin fusion protein was reconstituted with E. coli RNA polymerase core enzyme and tested in vitro. This chimeric holoenzyme specifically bound the spinach psbA and Chlamydomonas rrn16 promoters in gel mobility shift assays and exhibited specific transcription initiation from the same two promoters, providing evidence for the role of CrRpoD as a functional transcription factor. © Springer-Verlag 2006.
Scientific Publication
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