Characterization of Spiroplasma citri adhesion related protein SARP1, which contains a domain of a novel family designated sarpin.
Transmission of the plant pathogen Spiroplasma citri by its leafhopper vector, Circulifer tenellus, involves adherence to and invasion of insect host cells. The S. citri adhesion related protein P89 (SARP1) was purified by immunoprecipitation using anti-SARP1 monoclonal antibodies. The protein's N-terminal amino acid sequence was determined and used to design a degenerate oligonucleotide. The labeled oligonucleotide hybridized to a 3.5 kb MboI fragment from S. citri DNA, which was then cloned and sequenced. Additionally, a 1.9 kb RsaI fragment of S. citri DNA, partially overlapping the MboI fragment, was isolated and characterized. Sequence analysis of the two clones revealed four open reading frames. ORF1 (675 bp) encodes the C-terminal part of a Soj-like protein. ORFs 1 and 2 were separated from ORFs 3 and 4 by a putative transcription termination site, indicated by a hairpin structure. ORF3 encodes an amphiphilic 798 amino acid long protein with a cleavable signal peptide and a predicted transmembrane helix near the C-terminus. The mature protein of 85.96 kDa has a calculated pI value of 5.5 and has an N-terminal amino acid sequence consistent with that determined from the purified SARP1. At the N-terminus of this protein is a region consisting of six repeats, each 39-42 amino acids, a motif belonging to a previously unrecognized family of repeats found in a variety of bacterial proteins. The taxonomically spotty presence of this 'sarpin' domain and the relationship of the repeats to each other suggests a convergent evolution in multiple lineages.
Berg
M
M
Department of Entomology and Plant Pathology, Oklahoma State University, Stillwater, OK 74078, USA.
Melcher
U
U
Fletcher
J
J
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D013149:Spiroplasma / Q000235:genetics*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C440889:Spiroplasma citri adhesion related protein P89