Physical mapping and expression of gene families encoding the N-acetyl D-galactosamine adherence lectin of Entamoeba histolytica.
Adherence of the enteric protozoan parasite Entamoeba histolytica is mediated by an N-acetyl D-galactosamine (GalNAc)-specific lectin, a heterodimer of heavy (170 kDa) and light (35/31 kDa) subunits. The gene families encoding the lectin subunits were characterized using clamped homogeneous electric field (CHEF) gel electrophoresis in the strain HM1:IMSS. The heavy subunit was shown to be encoded by a family of five hgl genes, which were physically mapped to five distinct HindIII restriction fragments. The light subunit was shown to be encoded by a family of lgl genes located at six loci in the genome. Heavy and light subunit genes did not appear to be linked. Partial sequences of new members of the hgl and lgl gene families were obtained. Several different strains of E. histolytica were found to contain multiple hgl loci in their genomes. Expression of hgl and lgl genes in HM1:IMSS trophozoites was examined under different growth conditions using the reverse transcription-polymerase chain reaction (RT-PCR). mRNA transcripts were detected from three hgl genes and three lgl genes, with no significant differences between cultured amoebae and amoebae from liver abscesses. The complexity of GalNAc lectin gene expression observed suggests distinct biological functions for the products of the individual genes during pathogenesis.
Ramakrishnan
G
G
Department of Internal Medicine, University of Virginia, Charlottesville 22908, USA.
Ragland
B D
BD
Purdy
J E
JE
Mann
B J
BJ
D000116:Acetylgalactosamine / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D002448:Cell Adhesion / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D004748:Entamoeba histolytica / Q000378:metabolism*; D015870:Gene Expression / Q000235:genetics; D005838:Genotype; D037102:Lectins / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D016415:Sequence Alignment; D017421:Sequence Analysis
D001426:Bacterial Proteins; D017931:DNA Primers; D037102:Lectins; D000116:Acetylgalactosamine