Expression and function of a family of transmembrane kinases from the protozoan parasite Entamoeba histolytica.
The signaling proteome of Entamoeba histolytica is made of transmembrane kinases (TMKs) that are rarely found in unicellular eukaryotes. There are 90 TMK genes reported for E. histolytica, and these have been grouped into nine distinct families based on motifs present on both extracellular and kinase domains. Of these, the B1 family was chosen for further analysis. Genomic sequencing revealed the presence of 28 members belonging to this family. Genes corresponding to the majority of these were truncated and not considered for further analysis. Only five members were full length and contained both extracellular and cytosolic kinase domains. BLAST analysis revealed the presence of homologs of these B1 TMKs in the nonpathogenic Entamoeba dispar. However, the ligand binding domains of the orthologous B1 TMKs of the two species showed considerable divergence, indicating the possibility of a correlation with the pathogenic potential of the organism. Only two of the five full-length copies (B1.I.1 and B1.I.2) were expressed in E. histolytica under the culture conditions used. Antisera generated against the extracellular domain of B1.I.1 stained the cell surface, particularly the areas of contact between the trophozoites. Staining was also seen in the frontal and posterior regions of the motile amoeba. An amoebic cell line expressing a truncated version of the B1.I.1 that lacked the kinase domain was generated. Inducible expression of the truncated TMK resulted in a decrease in cellular proliferation and an increase in sensitivity to serum starvation. Our data indicate that the B1.I class of TMKs is involved in parasite proliferation.
Mehra
Alka
A
School of Life Sciences, Lab No. 117, Jawaharlal Nehru University, New Delhi-110067, India.
Fredrick
Jesse
J
Petri
William A
WA
Bhattacharya
Sudha
S
Bhattacharya
Alok
A
D000818:Animals; D002460:Cell Line; D019295:Computational Biology; D004748:Entamoeba histolytica / Q000201:enzymology* / Q000235:genetics*; D015870:Gene Expression; D017125:Genes, Protozoan; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D010641:Phenotype; D010770:Phosphotransferases / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*
D008565:Membrane Proteins; D015800:Protozoan Proteins; D010770:Phosphotransferases