DNA-binding specificity of the Lon protease alpha-domain from Brevibacillus thermoruber WR-249.
Lon protease has been well studied in many aspects; however, the DNA-binding specificity of Lon in prokaryotes has not been clearly identified. Here we examined the DNA-binding activity of Lon protease alpha-domains from Brevibacillus thermoruber (Bt), Bacillus subtilis (Bs), and Escherichia coli (Ec). MALDI-TOF mass spectroscopy showed that the alpha-domain from Bt-Lon binds to the duplex nucleotide sequence 5'-CTGTTAGCGGGC-3' (ms1) and protected it from DNase I digestion. Surface plasmon resonance showed that the Bt-Lon alpha-domain binds with ms1 double-stranded DNA tighter than Bs- and Ec-Lon alpha-domains, whereas the Bt-Lon alpha-domain has dramatically lower affinity for double-stranded DNA with 0 and 50% identity to the ms1 binding sequence. Our results indicated that Bt-Lon alpha-domain plays a critical role with ms1 sequence in the DNA-binding specificity.
Lin
Yu-Ching
YC
Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan.
Lee
Huai-Cheng
HC
Wang
Iren
I
Hsu
Chun-Hua
CH
Liao
Jiahn-Haur
JH
Lee
Alan Yueh-Luen
AY
Chen
Chinpan
C
Wu
Shih-Hsiung
SH
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology; D001419:Bacteria / Q000201:enzymology*; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000201:enzymology; D008969:Molecular Sequence Data; D049070:Protease La / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D004268:DNA-Binding Proteins; D004247:DNA; D049070:Protease La