Molecular cloning, structure and bait region splice variants of alpha2-macroglobulin from the soft tick Ornithodoros moubata.
The sequence of a alpha(2)-macroglobulin (alpha(2)M) from the soft tick Ornithodoros moubata (TAM) was determined by cloning and sequencing of overlapping polymerase chain reaction (PCR) and rapid amplification of cDNA ends PCR products. The TAM cDNA sequence is 4,944 bp long and contains one open reading frame coding for a protein precursor composed of 1,494 amino-acid residues, including a 24-residue signal sequence. The mature protein is cleaved into two subunits similarly to the C3 and C4 components of complement and fish alpha(2)Ms. Phylogeny analysis revealed that TAM is closely related to Limulus alpha(2)M and displays the highest similarity to the partial sequence of alpha(2)M from hard tick Ixodes scapularis. The comparison of conserved cysteine residues between TAM and human and Limulus alpha(2)Ms made it possible to predict the pattern of disulfide bridges and explain the atypical molecular arrangement of TAM. Four variants of the TAM bait region differing only in a short central segment were found; our data indicate that TAM exists as a single-copy gene in the tick genome and its bait region variants likely arise by alternative splicing. TAM is produced by tick hemocytes and it is also significantly expressed in salivary glands. TAM mRNA levels were shown to be up-regulated upon blood meal.
Saravanan
Thangamani
T
Institute of Parasitology, Academy of Sciences of the Czech Republic and Faculty of Biological Sciences, University of South Bohemia, Branisovská 31, 370 05 Ceské Budejovice, Czech Republic.
Weise
Christoph
C
Sojka
Daniel
D
Kopácek
Petr
P
D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D005247:Feeding Behavior; D005786:Gene Expression Regulation; D006023:Glycoproteins; D006434:Hemocytes / Q000737:chemistry; D019476:Insect Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000494:pharmacology; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D022821:RNA Splice Sites; D012469:Salivary Glands / Q000737:chemistry; D017422:Sequence Analysis, DNA; D013987:Ticks / Q000235:genetics*; D015854:Up-Regulation; D000511:alpha-Macroglobulins / Q000096:biosynthesis* / Q000235:genetics* / Q000494:pharmacology
D060829:Arthropod Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D006023:Glycoproteins; D019476:Insect Proteins; D022821:RNA Splice Sites; C404708:TAM macroglobulin protein, Ornithodoros moubata; D000511:alpha-Macroglobulins