Piriform spider silk sequences reveal unique repetitive elements.
Orb-weaving spider silk fibers are assembled from very large, highly repetitive proteins. The repeated segments contain, in turn, short, simple, and repetitive amino acid motifs that account for the physical and mechanical properties of the assembled fiber. Of the six orb-weaver silk fibroins, the piriform silk that makes the attachment discs, which lashes the joints of the web and attaches dragline silk to surfaces, has not been previously characterized. Piriform silk protein cDNAs were isolated from phage libraries of three species: A. trifasciata , N. clavipes , and N. cruentata . The deduced amino acid sequences from these genes revealed two new repetitive motifs: an alternating proline motif, where every other amino acid is proline, and a glutamine-rich motif of 6-8 amino acids. Similar to other spider silk proteins, the repeated segments are large (>200 amino acids) and highly homogenized within a species. There is also substantial sequence similarity across the genes from the three species, with particular conservation of the repetitive motifs. Northern blot analysis revealed that the mRNA is larger than 11 kb and is expressed exclusively in the piriform glands of the spider. Phylogenetic analysis of the C-terminal regions of the new proteins with published spidroins robustly shows that the piriform sequences form an ortholog group.
Perry
David J
DJ
Department of Molecular Biology, University of Wyoming, Laramie, Wyoming 821071-3944, United States, Laboratory of Molecular Biology, EMBRAPA Western Amazonia, Manaus, AM, Brazil, and Laboratory of Gene Transfer, Biotechnology Unit, EMBRAPA Genetic Resources and Biotechnology, Brasilia, DF, Brazil.
Bittencourt
Daniela
D
Siltberg-Liberles
Jessica
J
Rech
Elibio L
EL
Lewis
Randolph V
RV
D000595:Amino Acid Sequence; D000818:Animals; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment; D020086:Short Interspersed Nucleotide Elements / Q000235:genetics*; D047011:Silk / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D013045:Species Specificity; D013112:Spiders
D018076:DNA, Complementary; D012333:RNA, Messenger; D047011:Silk