Identification of Bombyx mori 14-3-3 orthologs and the interactor Hsp60.
The 14-3-3 protein family consists of evolutionarily conserved, acidic 30 kDa proteins composed of seven isoforms named beta, gamma, epsilon, zeta, eta, theta, and sigma in mammalian cells. The dimeric complex of 14-3-3 isoforms, acting as a molecular adaptor, plays a central role in regulation of neuronal function. Since aberrant expression of 14-3-3 is identified in the brains of Alzheimer disease and Parkinson disease, a convenient insect model, if it is available, is highly valuable for studying a pathological role of 14-3-3 in neurodegeneration. Here, we identified the silkworm Bombyx mori 14-3-3 orthologs, zeta and epsilon isoforms highly homologous in amino acid sequences to the human 14-3-3zeta and 14-3-3epsilon. By Western blot, the expression of zeta and epsilon isoforms was identified at substantial levels in the first instar larva, markedly upregulated in the second instar larva, and the highest levels were maintained in the late stage of larva, the pupa, and the adult. Furthermore, by protein overlay and immunoprecipitation, we identified Hsp60 as a 14-3-3-binding partner. The 14-3-3 proteins interacted with the N-terminal fragment of Hsp60. The 14-3-3zeta and epsilon isoforms, along with Hsp60, were expressed widely with overlapping distribution in larval and adult tissues, including brain, fat body, silk gland, Malpighian tube, midgut, ovary, testis, antenna, and pheromone gland. These observations suggest that a molecular adaptor 14-3-3 and a molecular chaperone Hsp60 cooperate to achieve a wide range of cellular functions in B. mori.
Tabunoki
Hiroko
H
Department of Bioinformatics and Molecular Neuropathology, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo 204-8588, Japan.
Shimada
Toru
T
Banno
Yutaka
Y
Sato
Ryoichi
R
Kajiwara
Hideyuki
H
Mita
Kazuei
K
Satoh
Jun-ichi
J
D048948:14-3-3 Proteins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D012831:Bombyx / Q000254:growth & development / Q000378:metabolism*; D018834:Chaperonin 60 / Q000378:metabolism*; D003001:Cloning, Molecular; D015180:Electrophoresis, Gel, Two-Dimensional; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D019476:Insect Proteins / Q000378:metabolism*; D007814:Larva; D008969:Molecular Sequence Data; D010802:Phylogeny; D011679:Pupa; D017386:Sequence Homology, Amino Acid
D048948:14-3-3 Proteins; D018834:Chaperonin 60; D019476:Insect Proteins