Stress and transcriptional regulation of tick ferritin HC.
We previously identified a partial Dermacentor variabilis cDNA encoding ferritin HC (HC) subunit homolog (DVFER) that was differentially upregulated in Rickettsia montanensis infected ticks (Mulenga et al., 2003a). We have used rapid amplification of cDNA ends to clone full-length DVFER cDNA and its apparent ortholog from the wood tick, D. andersoni (DAFER), both of which show high sequence similarity to vertebrate than insect ferritin. Both DVFER and DAFER contain the stem-loop structure of a putative iron responsive element in the 5' untranslated region (nucleotide positions, 16-42) and the feroxidase centre loop typical for vertebrate ferritin HC subunits. Quantitative Western and Northern blotting analyses of protein and RNA from unfed and partially fed whole tick as well as dissected tick tissues demonstrated that DVFER is constitutively and ubiquitously expressed. Based on densitometric analysis of detected protein and mRNA bands, DVFER is predominantly expressed in the midgut, and to a lesser extent in the salivary glands, ovary and fatbody. Sham treatment (mechanical injury) and Escherichia coli challenge of D. variabilis ticks stimulated statistically significant (approximately 1.5- and approximately 3.0-fold, respectively) increases in DVFER mRNA abundance over time point matched naive control ticks. These data suggest that DVFER mRNA is nonspecifically up regulated in response to mechanical injury or bacterial infection induced stress.
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD, USA. amvle001@umaryland.edu
A
A
Mulenga
J A
JA
Simser
K R
KR
Macaluso
A F
AF
Azad
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D003720:Densitometry; D004064:Digestive System / Q000378:metabolism; D004926:Escherichia coli; D005293:Ferritins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013987:Ticks / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D005293:Ferritins