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Identification of functional promoters in the msp2 expression loci of Anaplasma marginale and Anaplasma phagocytophilum.
Title | Identification of functional promoters in the msp2 expression loci of Anaplasma marginale and Anaplasma phagocytophilum. |
Publication Type | Journal Article |
Year of Publication | 2005 |
Authors | Barbet, AF, Agnes, JT, Moreland, AL, Lundgren, AM, A Alleman, R, Noh, SM, Brayton, KA, Munderloh, UG, Palmer, GH |
Journal | Gene |
Volume | 353 |
Issue | 1 |
Pagination | 89-97 |
Date Published | 2005 Jun 20 |
ISSN | 0378-1119 |
Keywords | Animals, Antigens, Bacterial, Bacterial Outer Membrane Proteins, Base Sequence, Cattle, Cell Line, Escherichia coli, Green Fluorescent Proteins, HL-60 Cells, Humans, Luminescent Measurements, Molecular Sequence Data, Promoter Regions, Genetic, Recombinant Fusion Proteins, Reverse Transcriptase Polymerase Chain Reaction, RNA, Messenger, Sequence Homology, Nucleic Acid, Transcription Initiation Site, Transcription, Genetic |
Abstract | Organisms in the family Anaplasmataceae are important tick-borne pathogens of livestock worldwide and cause recently emergent infections in humans. Despite their medical importance, very little is known about how these organisms regulate gene expression in the mammalian host, the tick vector, or during transition between the host and vector. However, it is clear that gene regulation, in addition to recombinatorial mechanisms, is essential for these small genome pathogens to adapt to distinctly different environments. In this study, we identify and establish the function of three promoter elements in the locus encoding major outer membrane protein expression sites in both Anaplasma marginale and Anaplasma phagocytophilum. Gene expression from this locus involves both classical and atypical polycistronic transcripts. The identified promoter elements have a structure similar to that defined in Escherichia coli and are functional in driving protein expression in a prokaryotic cell-free transcription and translation system and in recombinant E. coli. The two strongest promoters identified in vitro and with recombinant E. coli were also shown to be functional in A. marginale infected cells, as determined by quantification of downstream transcripts. The promoters in both A. marginale and A. phagocytophilum have similar structure and activity, supporting the conclusion that the two loci are syntenic with conservation of function. In addition, they share structural elements within the promoters that appear to be likely sites for regulation. These data enhance our understanding of how expression of these variable outer membrane proteins may be controlled in the key stages of tick-borne transmission and infection. |
DOI | 10.1016/j.gene.2005.03.036 |
Alternate Journal | Gene |
PubMed ID | 15935572 |
Grant List | K08 AI052412 / AI / NIAID NIH HHS / United States R01 AI042792 / AI / NIAID NIH HHS / United States R01 AI042792-06A1 / AI / NIAID NIH HHS / United States R01 AI44005 / AI / NIAID NIH HHS / United States R01 AI45580 / AI / NIAID NIH HHS / United States T32 GM008336 / GM / NIGMS NIH HHS / United States |