Heat shock and developmental expression of hsp83 in the filarial nematode Brugia pahangi.
hsp83 was cloned from the filarial nematode Brugia pahangi. The mRNA was constitutively expressed at 37 degrees C in life cycle stages that live in the mammalian host (microfilariae and adult worms). Heat shock resulted in only a minimal increase in levels of transcription. A genomic copy of hsp83 was isolated and was shown to contain 11 introns while sequencing of the 5' upstream region revealed several heat shock elements. Using a chloramphenicol acetyltransferase (CAT) reporter gene construct the expression of hsp83 from B. pahangi (Bp-hsp83) was studied by transfection of COS-7 cells. Similar to the expression pattern in the parasite, CAT activity was detected at 37 degrees C and was not influenced by heat shock. When the free-living nematode Caenorhabditis elegans was transfected with the same construct, CAT activity was not observed at normal growth temperatures (21 degrees C) or under moderate heat shock conditions (28 degrees C). However exposure to more severe heat shock (35 degrees C) resulted in an increase in CAT activity. These results suggest that Bp-hsp83 has a temperature threshold > or = 35 degrees C for expression.
Thompson
F J
FJ
Department of Veterinary Parasitology, University of Glasgow, Scotland, UK.
Cockroft
A C
AC
Wheatley
I
I
Britton
C
C
Devaney
E
E
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D017179:Brugia pahangi / Q000378:metabolism*; D019556:COS Cells; D003001:Cloning, Molecular; D018076:DNA, Complementary; D016054:DNA, Protozoan; D004797:Enzyme-Linked Immunosorbent Assay; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D018869:Heat-Shock Response; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D015854:Up-Regulation
D018076:DNA, Complementary; D016054:DNA, Protozoan; D006360:Heat-Shock Proteins; D012333:RNA, Messenger