TY - JOUR
T1 - Structural and genomic DNA analysis of a putative transcription factor SCO5550 from Streptomyces coelicolor A3(2)
T2 - Regulating the expression of gene sco5551 as a transcriptional activator with a novel dimer shape
AU - Hayashi, Takeshi
AU - Tanaka, Yoshikazu
AU - Sakai, Naoki
AU - Watanabe, Nobuhisa
AU - Tamura, Tomohiro
AU - Tanaka, Isao
AU - Yao, Min
N1 - Funding Information:
We thank Ms. T. Onodera and K. Hatakeyama for help with protein expression and purification. We also thank the staff of beamline BL44B2, Spring-8, Japan, for their kind help with data collection. This was supported by National Project on Protein Structural and Functional Analyses from the Ministry of Education, Culture, Sports, Science, and Technology of Japan .
PY - 2013/5/24
Y1 - 2013/5/24
N2 - SCO5550 from the model actinomycete Streptomyces coelicolor A3(2) was identified as a putative transcriptional regulator, and classified into the MerR family by sequence analysis. Recombined SCO5550 was successfully produced in Rhodococcus erythropolis, which can be used to stably express recombinant protein by optimizing the temperature over a wide range (4-35. °C). Crystal structure analysis showed that the dimerization domain (C-terminal domain) of SCO5550 has a novel fold and forms a new dimer shape, whereas the DNA-binding domain (N-terminal domain) is very similar to those of MerR family members. Such the new dimer form suggests that SCO5550 may define a new subfamily as a new member of the MerR family. Binding DNA sequence analysis of SCO5550 using the genomic systematic evolution of ligands by exponential enrichment (gSELEX) and electrophoretic mobility shift assay (EMSA) indicated that SCO5550 regulates the expression of the immediately upstream gene sco5551 encoding a putative protein, probably as a transcriptional activator.
AB - SCO5550 from the model actinomycete Streptomyces coelicolor A3(2) was identified as a putative transcriptional regulator, and classified into the MerR family by sequence analysis. Recombined SCO5550 was successfully produced in Rhodococcus erythropolis, which can be used to stably express recombinant protein by optimizing the temperature over a wide range (4-35. °C). Crystal structure analysis showed that the dimerization domain (C-terminal domain) of SCO5550 has a novel fold and forms a new dimer shape, whereas the DNA-binding domain (N-terminal domain) is very similar to those of MerR family members. Such the new dimer form suggests that SCO5550 may define a new subfamily as a new member of the MerR family. Binding DNA sequence analysis of SCO5550 using the genomic systematic evolution of ligands by exponential enrichment (gSELEX) and electrophoretic mobility shift assay (EMSA) indicated that SCO5550 regulates the expression of the immediately upstream gene sco5551 encoding a putative protein, probably as a transcriptional activator.
KW - GSELEX
KW - MerR family
KW - SCO5550
KW - Transcriptional activator
KW - X-ray crystallography
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U2 - 10.1016/j.bbrc.2013.04.017
DO - 10.1016/j.bbrc.2013.04.017
M3 - Article
C2 - 23618855
AN - SCOPUS:84878189923
VL - 435
SP - 28
EP - 33
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -