Identification and characterization of a novel toxin-antitoxin module from Bacillus anthracis

Shivangi Agarwal; Shivani Agarwal; Rakesh Bhatnagar

doi:10.1016/j.febslet.2007.03.051

Identification and characterization of a novel toxin-antitoxin module from Bacillus anthracis

Shivangi Agarwal, Shivani Agarwal, Rakesh Bhatnagar^*

^*Corresponding author for this work

Medicine, Hematology Oncology Division

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

Abstract

Comparative genome analysis of Bacillus anthracis revealed a pair of linked genes encoding pemK (K, killer protein) and pemI (I, inhibitory protein) homologous to pem loci of other organisms. Expression of PemK in Escherichia coli and Bacillus anthracis was bacteriostatic whereas the concomitant expression of PemI reversed the growth arrest. PemK expression effectively inhibited protein synthesis with no significant effect on DNA replication. Coexpression and interaction of these proteins confirmed it to be a Type II addiction module. Thermal denaturation analysis reflected poor conformational stability of PemI as compared to PemK. Circular dichroism analysis indicated that PemI contains twice the amount of β-sheets as PemK. Gel retardation assays demonstrated that PemI binds to its upstream DNA sequence. This study reports the first evidence of an active chromosome encoded toxin-antitoxin locus in B. anthracis.

Original language	English (US)
Pages (from-to)	1727-1734
Number of pages	8
Journal	FEBS Letters
Volume	581
Issue number	9
DOIs	https://doi.org/10.1016/j.febslet.2007.03.051
State	Published - May 1 2007

Keywords

Addiction module
Bacillus anthracis
Plasmid inheritance
Post segregational killing
Toxin-antitoxin locus

ASJC Scopus subject areas

Genetics
Molecular Biology
Biophysics
Structural Biology
Biochemistry
Cell Biology

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title = "Identification and characterization of a novel toxin-antitoxin module from Bacillus anthracis",

abstract = "Comparative genome analysis of Bacillus anthracis revealed a pair of linked genes encoding pemK (K, killer protein) and pemI (I, inhibitory protein) homologous to pem loci of other organisms. Expression of PemK in Escherichia coli and Bacillus anthracis was bacteriostatic whereas the concomitant expression of PemI reversed the growth arrest. PemK expression effectively inhibited protein synthesis with no significant effect on DNA replication. Coexpression and interaction of these proteins confirmed it to be a Type II addiction module. Thermal denaturation analysis reflected poor conformational stability of PemI as compared to PemK. Circular dichroism analysis indicated that PemI contains twice the amount of β-sheets as PemK. Gel retardation assays demonstrated that PemI binds to its upstream DNA sequence. This study reports the first evidence of an active chromosome encoded toxin-antitoxin locus in B. anthracis.",

keywords = "Addiction module, Bacillus anthracis, Plasmid inheritance, Post segregational killing, Toxin-antitoxin locus",

author = "Shivangi Agarwal and Shivani Agarwal and Rakesh Bhatnagar",

note = "Funding Information: We acknowledge financial support from the Department of Biotechnology, Government of India. Special thanks go to Dr. N. Banerjee, ICGEB, New Delhi, India and Dr. A. Kulshrestha, Nanyang Technological University, Singapore, for constructive and valuable discussions.",

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doi = "10.1016/j.febslet.2007.03.051",

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AU - Agarwal, Shivani

AU - Bhatnagar, Rakesh

N1 - Funding Information: We acknowledge financial support from the Department of Biotechnology, Government of India. Special thanks go to Dr. N. Banerjee, ICGEB, New Delhi, India and Dr. A. Kulshrestha, Nanyang Technological University, Singapore, for constructive and valuable discussions.

PY - 2007/5/1

Y1 - 2007/5/1

N2 - Comparative genome analysis of Bacillus anthracis revealed a pair of linked genes encoding pemK (K, killer protein) and pemI (I, inhibitory protein) homologous to pem loci of other organisms. Expression of PemK in Escherichia coli and Bacillus anthracis was bacteriostatic whereas the concomitant expression of PemI reversed the growth arrest. PemK expression effectively inhibited protein synthesis with no significant effect on DNA replication. Coexpression and interaction of these proteins confirmed it to be a Type II addiction module. Thermal denaturation analysis reflected poor conformational stability of PemI as compared to PemK. Circular dichroism analysis indicated that PemI contains twice the amount of β-sheets as PemK. Gel retardation assays demonstrated that PemI binds to its upstream DNA sequence. This study reports the first evidence of an active chromosome encoded toxin-antitoxin locus in B. anthracis.

AB - Comparative genome analysis of Bacillus anthracis revealed a pair of linked genes encoding pemK (K, killer protein) and pemI (I, inhibitory protein) homologous to pem loci of other organisms. Expression of PemK in Escherichia coli and Bacillus anthracis was bacteriostatic whereas the concomitant expression of PemI reversed the growth arrest. PemK expression effectively inhibited protein synthesis with no significant effect on DNA replication. Coexpression and interaction of these proteins confirmed it to be a Type II addiction module. Thermal denaturation analysis reflected poor conformational stability of PemI as compared to PemK. Circular dichroism analysis indicated that PemI contains twice the amount of β-sheets as PemK. Gel retardation assays demonstrated that PemI binds to its upstream DNA sequence. This study reports the first evidence of an active chromosome encoded toxin-antitoxin locus in B. anthracis.

KW - Addiction module

KW - Bacillus anthracis

KW - Plasmid inheritance

KW - Post segregational killing

KW - Toxin-antitoxin locus

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Identification and characterization of a novel toxin-antitoxin module from Bacillus anthracis

Abstract

Keywords

ASJC Scopus subject areas

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