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In vivo splicing and functional characterization of Mycobacterium leprae RecA
K Frischkorn1, B Springer, E C Böttger
1Medizinische Hochschule Hannover, Institut für Medizinische Mikrobiologie, 30625 Hannover, Germany.
Journal of Bacteriology
|June 15, 2000
Summary
Mycobacterium leprae RecA protein undergoes functional splicing in Mycobacterium smegmatis, unlike in E. coli. This splicing confers resistance to DNA damage and enables homologous recombination.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- RecA proteins are crucial for DNA repair and recombination.
- Inteins are protein segments that can excise themselves.
- Mycobacterium tuberculosis and Mycobacterium leprae RecA proteins possess inteins.
Purpose of the Study:
- To investigate the functional splicing of Mycobacterium leprae RecA protein.
- To determine if Mycobacterium leprae RecA can be spliced in a heterologous host.
- To assess the functional consequences of M. leprae RecA splicing.
Main Methods:
- Expression of Mycobacterium leprae RecA in Escherichia coli and Mycobacterium smegmatis.
- Assessing RecA splicing efficiency in different bacterial hosts.
- Evaluating DNA-damaging agent resistance and homologous recombination assays.
Main Results:
- Mycobacterium leprae RecA did not splice in Escherichia coli.
- Functional splicing of Mycobacterium leprae RecA was observed in Mycobacterium smegmatis.
- Spliced M. leprae RecA conferred resistance to DNA-damaging agents.
- Spliced M. leprae RecA facilitated homologous recombination.
Conclusions:
- Mycobacterium smegmatis is a suitable host for M. leprae RecA intein splicing.
- Functional M. leprae RecA protein is produced via intein splicing in M. smegmatis.
- Intein-mediated splicing of M. leprae RecA is essential for its DNA repair and recombination functions.