T cell receptor beta chain from sea bream (Sparus aurata): molecular cloning, expression and modelling of the complexes with MHC class I.
The T cell receptor is a fundamental mediator of the adaptive immune responses, since TR alphabeta on T cells recognize foreign structures (peptides derived from processed antigens) bound to the major histocompatibility complex (MHC) on APC cells. In the present study, we report the cloning of six TRB chains cDNA sequences from gilthead sea bream (Sparus aurata), a fish of high economical impact in South Mediterranean aquaculture. The V-BETA domains have the canonical features of known teleost and mammalian TR V-BETA domains and have been divided in four different subgroups. A multiple alignment of the six sea bream TRB chains with other known TRB sequences was assembled and showed the conservation of the four cysteine residues involved in disulphide bonds and of some amino acids with an important role in the assembly and signalling of the TR alphabeta/CD3 complex. Real-time PCR analysis was used to investigate TRB basal expression, that was maximum in the thymus followed by gut, and TRB in vitro expression after stimulation with LPS or PHA-L at 4 and 24h (only the 4h stimulation with LPS gave a significant effect). Moreover, the 3D structures of sea bream TRB chains and MHC-I were predicted by homology modelling with the final aim to investigate the interaction surface in the V-BETA/MHC-I complexes.
Dipartimento di Scienze Ambientali, Università della Tuscia, Largo dell'Università s.n.c., I-01100 Viterbo, Italy.
Elisa
E
Randelli
Viviana
V
Scala
Daniela
D
Casani
Susan
S
Costantini
Angelo
A
Facchiano
Massimo
M
Mazzini
Giuseppe
G
Scapigliati
Francesco
F
Buonocore
D000595:Amino Acid Sequence; D000818:Animals; D002999:Clone Cells; D003001:Cloning, Molecular; D003198:Computer Simulation; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D015395:Histocompatibility Antigens Class I / Q000737:chemistry*; D006860:Hydrogen Bonding; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D021541:Sea Bream / Q000235:genetics*; D016415:Sequence Alignment; D013816:Thermodynamics
D015395:Histocompatibility Antigens Class I; D016693:Receptors, Antigen, T-Cell, alpha-beta