Molecular characterization and real-time PCR transcriptional analysis of Dictyocaulus viviparus major sperm proteins.
Major sperm proteins (MSPs) represent a protein family occurring in nematodes only. Identification of the 3' and 5' untranslated region (UTR) completed the so far partial msp complementary DNA sequences of the bovine lungworm Dictyocaulus viviparus. The full-length transcript contains sequence tracts consistent with the Kozak and polyadenylation consensus sequence. On genomic level, three full-length sequences differing in three nucleotides were determined containing a 65-bp phase zero intron. Conceptual translation inferred two MSP isoforms due to one substitution within the 126-amino acid polypeptide. Bioinformatic analysis predicted that bovine lungworm MSP folds into an immunoglobulin-like seven-stranded beta sandwich as known for Caenorhabditis elegans and Ascaris suum. Furthermore, bovine lungworm MSP is confidentially predicted to be N-terminal-acetylated and secreted via a non-classical pathway. Quantitative real-time polymerase chain reaction analysis using ten developmental lungworm stages showed that msp is transcribed mainly in adult male parasites and in some degree in hypobiotic L5. However, marginal msp transcription was detectable in all of the investigated developmental lungworm stages.
Institute for Parasitology, University of Veterinary Medicine Hannover, Buenteweg 17, 30559, Hannover, Germany. christina.strube@tiho-hannover.de
Christina
C
Strube
Sandra
S
Buschbaum
Thomas
T
Schnieder
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002417:Cattle; D019295:Computational Biology; D018076:DNA, Complementary; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D004021:Dictyocaulus / Q000737:chemistry* / Q000235:genetics*; D020869:Gene Expression Profiling; D015801:Helminth Proteins / Q000096:biosynthesis* / Q000235:genetics*; D007438:Introns; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D017510:Protein Folding; D020033:Protein Isoforms / Q000235:genetics; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D020033:Protein Isoforms; C050602:major sperm protein, nematode