Characterization of the BH3 protein Bmf in Gallus gallus: identification of a novel chicken-specific isoform.
Bmf is a proapoptotic member of the BH3-only subgroup of Bcl-2 family proteins, which is associated to myosin V motors by binding to the dynein light chain 2 (DLC2). It acts as a sentinel detecting intracellular damages on the main cytoskeletal structures. The cloning and characterization of the chicken (Gallus gallus) Bmf cDNA and splicing variant is described in this report. The Bmf cDNA was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) using oligonucleotide primers derived from in silico sequences. The chicken Bmf cDNA encodes a protein of 193 amino acids, showing homology to mammalian Bmf proteins. A splicing variant of the chicken Bmf (Bmf(S), short isoform of Bmf) coding a protein of 118 amino acids was also identified. This is the first Bmf isoform identified so far which lacks the DLC2-binding domain although retaining the BH3 domain. Both chicken Bmf isoforms induced apoptosis 24 h after transfection in MCF7 and HeLa cell lines, but chicken Bmf(S) exhibits a higher proapoptotic activity. In addition, mRNA expression analysis showed that chicken Bmf transcription is ubiquitous in all embryo developmental stages, suggesting a role for this protein in the control of the development process.
Gómez-Esquer
Francisco
F
Departamento de Ciencias de la Salud III, Universidad Rey Juan Carlos, Avenida de Atenas S/N, Alcorcón, Madrid, Spain.
Palomar
Ma Angustias
MA
Rivas
Inés
I
Delcán
José
J
Linares
Rafael
R
Díaz-Gil
Gema
G
D048868:Adaptor Proteins, Signal Transducing / Q000032:analysis / Q000145:classification / Q000235:genetics*; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D001483:Base Sequence; D002645:Chickens / Q000235:genetics* / Q000254:growth & development; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D020033:Protein Isoforms / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D048868:Adaptor Proteins, Signal Transducing; D018076:DNA, Complementary; D020033:Protein Isoforms; D012333:RNA, Messenger