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Conserved inserts in the Hsp60 (GroEL) and Hsp70 (DnaK) proteins are essential for cellular growth

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Abstract

The Hsp60 and Hsp70 chaperones contain a number of conserved inserts that are restricted to particular phyla of bacteria. A one aa insert in the E. coli GroEL and a 21–23 insert in the DnaK proteins are specific for most Gram-negative bacteria. Two other inserts in DnaK are limited to certain groups of proteobacteria. The requirement of these inserts for cellular growth was examined by carrying out complementation studies with temperature-sensitive (T s) mutants of E. coli groEL or dnaK. Our results demonstrate that deletion or most changes in these inserts completely abolished the complementation ability of the mutant proteins. Studies with GroEL and DnaK from some other species that either lacked or contained these inserts also indicated that these inserts are essential for growth of E. coli. The DnaK from some bacteria contains a two aa insert that is not found in E. coli. Introduction of this insert into the E. coli DnaK also led to its inactivation, indicating that these inserts are specific for different groups. We postulate that these conserved inserts that are localized in loop regions on protein surfaces, are involved in some ancillary functions that are essential for the groups of bacteria where they are found.

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Acknowledgments

We are grateful to Drs. Costa Georgopoulos and Debbie Ang for sending us the T s mutant strains for groEL and dnaK that were used in these studies. We also thank Drs. Lori Burrows and Dr. T. Finan for the DNA from P. aeruginosa and S. meliloti, respectively. This work was supported by a research grant from the Canadian Institute of Health Research.

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  1. Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, L8N 3Z5, Canada

    Bhag Singh & Radhey S. Gupta

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  1. Bhag Singh
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  2. Radhey S. Gupta
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Correspondence to Radhey S. Gupta.

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Communicated by D. Andersson.

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Singh, B., Gupta, R.S. Conserved inserts in the Hsp60 (GroEL) and Hsp70 (DnaK) proteins are essential for cellular growth. Mol Genet Genomics 281, 361–373 (2009). https://doi.org/10.1007/s00438-008-0417-3

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