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Two highly thermostable paralogous single-stranded DNA-binding proteins from Thermoanaerobacter tengcongensis
Abstract
The thermophilic bacterium Thermoanaerobacter tengcongensis has two single-stranded DNA-binding (SSB) proteins, designated TteSSB2 and TteSSB3. In a SSB complementation assay in Escherichia coli, only TteSSB3 took over the in vivo function of EcoSSB. We have cloned the ssb genes obtained by PCR and have developed E. coli overexpression systems. The TteSSB2 and TteSSB3 consist of 153 and 150 amino acids with a calculated molecular mass of 17.29 and 16.96 kDa, respectively. They are the smallest known bacterial SSB proteins. The homology between amino acid sequences of these proteins is 40% identity and 53% similarity. They are functional as homotetramers, with each monomer encoding one single-stranded DNA binding domain (OB-fold). In fluorescence titrations with poly(dT), both proteins bind single-stranded DNA with a binding site size of about 40 nt per homotetramer. Thermostability with half-life of about 30 s at 95°C makes TteSSB3 similar to the known SSB of Thermus aquaticus (TaqSSB). The TteSSB2 was fully active even after 6 h incubation at 100°C. Here, we show for the first time paralogous thermostable homotetrameric SSBs, which could be an attractive alternative for known homodimeric thermostable SSB proteins in their applications for molecular biology methods and analytical purposes.
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- Category:
- Articles
- Type:
- artykuł w czasopiśmie z listy filadelfijskiej
- Published in:
-
ARCHIVES OF MICROBIOLOGY
no. 190,
pages 79 - 87,
ISSN: 0302-8933 - Language:
- English
- Publication year:
- 2008
- Bibliographic description:
- Olszewski M., Mickiewicz M., Kur J.: Two highly thermostable paralogous single-stranded DNA-binding proteins from Thermoanaerobacter tengcongensis// ARCHIVES OF MICROBIOLOGY. -Vol. 190., nr. nr 1 (2008), s.79-87
- DOI:
- Digital Object Identifier (open in new tab) 10.1007/s00203-008-0366-6
- Verified by:
- Gdańsk University of Technology
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