Cloning and characterization of the target protein subunit lst8 of rapamycin in Apostichopus japonicus.

Autor: Yue Z; State Key Laboratory for Quality and Safety of Agro-products, Ningbo University, Ningbo, 315211, PR China., Lv Z; State Key Laboratory for Quality and Safety of Agro-products, Ningbo University, Ningbo, 315211, PR China., Shao Y; State Key Laboratory for Quality and Safety of Agro-products, Ningbo University, Ningbo, 315211, PR China., Zhang W; State Key Laboratory for Quality and Safety of Agro-products, Ningbo University, Ningbo, 315211, PR China., Zhao X; State Key Laboratory for Quality and Safety of Agro-products, Ningbo University, Ningbo, 315211, PR China., Guo M; State Key Laboratory for Quality and Safety of Agro-products, Ningbo University, Ningbo, 315211, PR China., Li C; State Key Laboratory for Quality and Safety of Agro-products, Ningbo University, Ningbo, 315211, PR China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, PR China. Electronic address: lichenghua@nbu.edu.cn.
Jazyk: angličtina
Zdroj: Fish & shellfish immunology [Fish Shellfish Immunol] 2019 Sep; Vol. 92, pp. 460-468. Date of Electronic Publication: 2019 Jun 21.
DOI: 10.1016/j.fsi.2019.06.038
Abstrakt: Autophagy plays an important role in the immune defense systems of vertebrates through the interaction between the lethal with SEC13 protein 8 (lst8) and the mechanistic target of rapamycin. In the present study, a novel invertebrate lst8 homologue is identified from Apostichopus japonicus (designated as Ajlst8) via polymerase chain reaction. The full-length complementary DNA of Ajlst8 comprises a 5'-untranslated region (UTR) of 78 base pair (bp), a 3'-UTR of 479 bp, and a putative open reading frame of 951 bp; hence, 316 amino acids are encoded. Structural analysis shows that the deduced amino acid of Ajlst8 shares six typical WD40 domains (28 aa-248 aa). Spatial expression analysis indicates that Ajlst8 is ubiquitously expressed in all the examined tissues, with a larger magnitude in coelomocytes. Vibrio splendidus infection in vivo and lipopolysaccharide exposure in vitro can significantly upregulate the messenger RNA expression of Ajlst8 by 2.39-fold and 1.93-fold compared with the control group, respectively. LPS exposure could also significantly induced the protein level of Ajlst8 to 2.38-fold and the autophagy level was markedly increased by 3.08-fold under same condition. The RNA interference of Ajlst8 in primary coelomocytes also reduces the relative expression of autophagy with a 0.71-fold decrease in the ratio of LC3-II/LC3-I compared with that in the control group. These results indicate that Ajlst8 is a novel immune regulator that may be involved in the antibacterial response process of sea cucumber by regulating autophagy.
(Copyright © 2019 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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