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Identification and Characterization of T heileria annulata Heat‐Shock Protein 90 (HSP90) Isoforms
Author(s) -
Mohammed S. B.,
Bakheit M. A.,
Ernst M.,
Ahmed J. S.,
Seitzer U.
Publication year - 2013
Publication title -
transboundary and emerging diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.392
H-Index - 63
eISSN - 1865-1682
pISSN - 1865-1674
DOI - 10.1111/tbed.12150
Subject(s) - geldanamycin , heat shock protein , biology , theileria , hsp90 , western blot , microbiology and biotechnology , apoptosis , hsp70 , gene isoform , blot , recombinant dna , gene , biochemistry , parasite hosting , world wide web , computer science
Summary Heat‐shock proteins (HSPs) refer to a group of proteins whose synthesis is enhanced upon sudden increase in temperature or exposure to a variety of other stressors. In this study, Theileria annulata ( T. annulata ) HSP90 was identified and characterized as a first step to understand the function of this molecule in T. annulata ‐infected cells. Our results indicated the existence in the genome of T. annulata of two HSP90 genes: one located in chromosome one ( Ta HSP90‐Chr1) and the other in chromosome four ( Ta HSP90‐Chr4). The amino acid alignment between the two isoforms has shown identity and similarity values of 23.52% and 30.26%, respectively. Theileria annulata recombinant HSP90 proteins were expressed using a bacterial expression system and could be recognized in Western blots by rabbit anti‐serum raised against an antigenic peptide derived from a unique sequence of Ta HSP90‐Chr1. On the other hand, bovine HSP90 was detected in T. annulata‐ infected cells using Western blot and immunocytostaining. To demonstrate the effect of the inhibition of HSP90 on the survival of T. annulata ‐infected cells, Geldanamycin (GA), a specific inhibitor for HSP90, was used. Upon GA treatment, p53 was observed to translocate into the host cell nucleus, a phenomenon that occurs in cells undergoing apoptosis. Using flowcytometry, a significant increase ( P = 0.028) in cell death (%) was observed in T. annulata‐ infected cells treated with two different GA concentrations, 0.5 and 1 μ m, and incubated for 24, 48 and 72 h.