The hypothetical protein 'All4779', and not the annotated 'Alr0088' and 'Alr7579' proteins, is the major typical single-stranded DNA binding protein of the cyanobacterium, Anabaena sp. PCC7120.
Single-stranded DNA binding (SSB) proteins are essential for all DNA-dependent cellular processes. Typical SSB proteins have an N-terminal Oligonucleotide-Binding (OB) fold, a Proline/Glycine rich region, followed by a C-terminal acidic tail. In the genome of the heterocystous nitrogen-fixing cyanobacterium, Anabaena sp. strain PCC7120, alr0088 and alr7579 are annotated as coding for SSB, but are truncated and have only the OB-fold. In silico analysis of whole genome of Anabaena sp. strain PCC7120 revealed the presence of another ORF 'all4779', annotated as a hypothetical protein, but having an N-terminal OB-fold, a P/G-rich region and a C-terminal acidic tail. Biochemical characterisation of all three purified recombinant proteins revealed that they exist either as monomer or dimer and bind ssDNA, but differently. The All4779 bound ssDNA in two binding modes i.e. (All4779)35 and (All4779)66 depending on salt concentration and with a binding affinity similar to that of Escherichia coli SSB. On the other hand, Alr0088 bound in a single binding mode of 50-mer and Alr7579 only to large stretches of ssDNA, suggesting that All4779, in all likelihood, is the major typical bacterial SSB in Anabaena. Overexpression of All4779 in Anabaena sp. strain PCC7120 led to enhancement of tolerance to DNA-damaging stresses, such as γ-rays, UV-irradiation, desiccation and mitomycinC exposure. The tolerance appears to be a consequence of reduced DNA damage or efficient DNA repair due to increased availability of All4779. The ORF all4779 is proposed to be re-annotated as Anabaena ssb gene.
Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, India.
Anurag
A
Kirti
Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, India.
Hema
H
Rajaram
Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, India.
Shree Kumar
SK
Apte
D000595:Amino Acid Sequence; D017033:Anabaena / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004260:DNA Repair / Q000235:genetics; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D030781:Organisms, Genetically Modified; D055049:Salt Tolerance / Q000235:genetics; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins