Cloning, expression and characterization of the bovine p65 subunit of NFkappaB.
The full length coding sequence of the cattle transcription factor p65 was isolated and cloned. The cloned bovine p65 was expressed in mammalian cells, and it induced the NF-kappaB-specific luciferase reporter gene expression. Using gel retardation techniques, we demonstrated that the cloned bovine p65 bound to the consensus kappaB sequence. The comparison of the bovine p65 with its human and mouse orthologues at amino acid level showed high homology in both the DNA-binding domain, known as Rel homology domain (RHD) and the transactivation domain (TAD). The phylogenetic analysis at DNA level provided a new insight in the evolution of the NF-kappaB family, and it could resolve the topology of the mammalian p65molecules. Although, the RHD was conserved in vertebrates, the TAD sequences deviated from each other, and showed faster molecular evolution than RHD sequences, which may indirectly result in the modification of NF-kappaB immune functions.
Doleschall
Márton
M
Department of Physiology and Biochemistry, Faculty of Veterinary Science, Szent István University, István u. 2. H-1078, Budapest, Hungary.
Mayer
Balázs
B
Cervenak
Judit
J
Cervenak
László
L
Kacskovics
Imre
I
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D017385:Sequence Homology; D051996:Transcription Factor RelA / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000378:metabolism*
D018076:DNA, Complementary; D051996:Transcription Factor RelA