Beta-keratins of turtle shell are glycine-proline-tyrosine rich proteins similar to those of crocodilians and birds.
This study presents, for the first time, sequences of five beta-keratin cDNAs from turtle epidermis obtained by means of 5'- and 3'-rapid amplification of cDNA ends (RACE) analyses. The deduced amino acid sequences correspond to distinct glycine-proline-serine-tyrosine rich proteins containing 122-174 amino acids. In situ hybridization shows that beta-keratin mRNAs are expressed in cells of the differentiating beta-layers of the shell scutes. Southern blotting analysis reveals that turtle beta-keratins belong to a well-conserved multigene family. This result was confirmed by the amplification and sequencing of 13 genomic fragments corresponding to beta-keratin genes. Like snake, crocodile and avian beta-keratin genes, turtle beta-keratins contain an intron that interrupts the 5'-untranslated region. The length of the intron is variable, ranging from 0.35 to 1.00 kb. One of the sequences obtained from genomic amplifications corresponds to one of the five sequences obtained from cDNA cloning; thus, sequences of a total of 17 turtle beta-keratins were determined in the present study. The predicted molecular weight of the 17 different deduced proteins range from 11.9 to 17.0 kDa with a predicted isoelectric point of 6.8-8.4; therefore, they are neutral to basic proteins. A central region rich in proline and with beta-strand conformation shows high conservation with other reptilian and avian beta-keratins, and it is likely involved in their polymerization. Glycine repeat regions, often containing tyrosine, are localized toward the C-terminus. Phylogenetic analysis shows that turtle beta-keratins are more similar to crocodilian and avian beta-keratins than to those of lizards and snakes.
Dalla Valle
Luisa
L
Dipartimento di Biologia, Università di Padova, Italy.
Nardi
Alessia
A
Toni
Mattia
M
Emera
Deena
D
Alibardi
Lorenzo
L
D000489:Alligators and Crocodiles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001717:Birds; D015152:Blotting, Northern / Q000379:methods; D015139:Blotting, Southern / Q000379:methods; D003001:Cloning, Molecular; D015870:Gene Expression; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000379:methods; D008969:Molecular Sequence Data; D010802:Phylogeny; D011487:Protein Conformation; D030162:Reptilian Proteins / Q000032:analysis / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012867:Skin / Q000378:metabolism*; D014426:Turtles / Q000235:genetics* / Q000378:metabolism; D053800:beta-Keratins / Q000032:analysis / Q000235:genetics*
D030162:Reptilian Proteins; D053800:beta-Keratins