Myosins of Babesia bovis: molecular characterisation, erythrocyte invasion, and phylogeny.
Using degenerate primers, three putative myosin sequences were amplified from Australian isolates of Babesa bovis and confirmed as myosins (termed Bbmyo-A, Bbmyo-B, and Bbmyo-C) from in vitro cultures of the W strain of B. bovis. Comprehensive analysis of 15 apicomplexan myosins suggests that members of Class XIV be defined as those with greater than 35% myosin head sequence identity and that these be further subclassed into groups bearing above 50-60% identity. Bbmyo-A protein bears a strong similarity with other apicomplexan myosin-A type proteins (subclass XIVa), the Bbmyo-B myosin head protein sequence exhibits low identity (35-39%) with all members of Class XIV, and 5'-sequence of Bbmyo-C shows strong identity (60%) with P. falciparum myosin-C protein. Domain analysis revealed five divergent IQ domains within the neck of Pfmyo-C, and a myosin-N terminal domain as well as a classical IQ sequence unusually located within the head converter domain of Bbmyo-B. A cross-reacting antibody directed against P. falciparum myosin-A (Pfmyo-A) revealed a zone of approximately 85 kDa in immunoblots prepared with B. bovis total protein, and immunofluorescence inferred stage-specific myosin-A expression since only 25% of infected erythrocytes with mostly paired B. bovis were immuno-positive. Multiplication of B. bovis in in vitro culture was inhibited by myosin- and actin-binding drugs at concentrations lower than those that inhibit P. falciparum. This study identifies and classifies three myosin genes and an actin gene in B. bovis, and provides the first evidence for the participation of an actomyosin-based motor in erythrocyte invasion in this species of apicomplexan parasite.
Queensland Department of Primary Industries, Moorooka, 4105, Queensland, Australia. Ala.Lew@dpi.qld.gov.au
A E
AE
Lew
A R
AR
Dluzewski
A M
AM
Johnson
J C
JC
Pinder
D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D016782:Apicomplexa / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016793:Babesia bovis / Q000235:genetics / Q000502:physiology*; D001483:Base Sequence; D002417:Cattle; D004912:Erythrocytes / Q000469:parasitology*; D017125:Genes, Protozoan; D006801:Humans; D008969:Molecular Sequence Data; D005810:Multigene Family; D009218:Myosins / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D015800:Protozoan Proteins / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D000199:Actins; D015800:Protozoan Proteins; D009218:Myosins