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Integration into the phage attachment site, attB, impairs multicellular differentiation in Stigmatella aurantiaca. J Bacteriol 2006 Mar;188(5):1701-9

Date

02/18/2006

Pubmed ID

16484181

Pubmed Central ID

PMC1426541

DOI

10.1128/JB.188.5.1701-1709.2006

Scopus ID

2-s2.0-33644783109   5 Citations

Abstract

Stigmatella aurantiaca displays a complex developmental life cycle in response to starvation conditions that results in the formation of tree-like fruiting bodies capable of producing spores. The phage Mx8, first isolated from the close relative Myxococcus xanthus, is unable to infect S. aurantiaca cells and integrate into the genome. However, plasmids containing Mx8 fragments encoding the integrase and attP are able to integrate at the attB locus in the S. aurantiaca genome by site-specific recombination. After recombination between attP and attB, the S. aurantiaca cells were incapable of building normal fruiting bodies but formed clumps and fungus-like structures characteristic of intermediate stages of development displayed by the wild type. We identified two tRNA genes, trnD and trnV, encoding tRNA(Asp) and tRNA(Val), respectively, composing an operon at the attB locus of S. aurantiaca. Integration of attP-containing plasmids resulted in the incorporation of the t(Mx8) terminator sequence, in addition to a short sequence of Mx8 DNA downstream of trnD. The integrant was unable to process the trnD transcript at the normal 3' processing site and displayed a lower level of expression of the trnVD operon. In addition, several developmentally regulated proteins were no longer produced in mutants following insertion at the attB locus. We hypothesize that the integration of the t(Mx8) terminator sequence results in reduced levels of mature tRNA(Asp) and tRNA(Val) and that altered protein production during development is thereby responsible for the observed phenotype. The trnVD locus thus defines a new developmental checkpoint for Stigmatella aurantiaca.

Author List

Müller S, Shen H, Hofmann D, Schairer HU, Kirby JR

Author

John Kirby PhD Chair, Center Associate Director, Professor in the Microbiology and Immunology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Attachment Sites, Microbiological
Bacterial Proteins
Bacteriophages
Base Sequence
Gene Expression Regulation, Bacterial
Genes, Bacterial
Genetic Complementation Test
Integrases
Molecular Sequence Data
Mutagenesis, Site-Directed
Operon
Plasmids
RNA, Transfer, Asp
RNA, Transfer, Val
Sequence Alignment
Spores, Bacterial
Stigmatella aurantiaca
Viral Proteins