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1.
Fish Shellfish Immunol ; 93: 743-751, 2019 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-31408731

RESUMO

White shrimp Litopenaeus vannamei are widely cultured in the world and white spot syndrome virus (WSSV) led to huge economic losses in the shrimp industry every year. In the present study, miRNAs involved in the response of shrimp L. vannamei to WSSV infection were obtained through the Illumina HiSeq 2500 high-throughput next-generation sequencing technique. A total number of 7 known miRNAs and 54 putative novel miRNAs were obtained. Among them, 14 DEMs were identified in the shrimp infected with WSSV. The putative target genes of these DEMs were related to host immune response or signaling pathways, indicating the importance of miRNAs in shrimp against WSSV infection. The results will provide information for further research on shrimp response to virus infection and contribute to the development of new strategies for effective protection against WSSV infections.

2.
Fish Shellfish Immunol ; 86: 1009-1018, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30586633

RESUMO

Autophagy plays a vital role in innate and adaptive immunity against invading microorganisms, such as virus and bacteria. However, the mechanism underlying autophagy in shrimp is still limited. In our study, we challenged white shrimp L. vannamei with rapamycin to induce autophagy and employed Solexa/Illumina high-throughput RNA-seq method to examine the differences of transcriptome from gills of shrimps treated with or without rapamycin. More than 22.64 Gb raw data were produced, which were assembled into 62, 503 unigenes, with 14,126 unigenes over 1 kb in length. We then performed differential expression analysis and identified a total of 3050 differentially expressed genes (DEGs). Among them, 1456 were upregulated and 1594 were downregulated. We further annotated DEGs by matching against non-redundant protein sequence (Nr), Swiss-Prot, Kyoto Encyclopedia of Genes and Genomes (KEGG), Clusters of Orthologous Groups of proteins (COG), euKaryotic Orthologous Groups (KOG), Gene ontology (GO), and Pfam databases. The assembled and annotated DEGs will facilitate our understanding of the molecular mechanism underlying autophagy and promote the studies on the role of autophagy in innate immunity of L. vannamei and other crustaceans.


Assuntos
Autofagia/imunologia , Penaeidae/genética , Penaeidae/imunologia , Transcriptoma , Animais , Autofagia/efeitos dos fármacos , Perfilação da Expressão Gênica , Brânquias/imunologia , Sequenciamento de Nucleotídeos em Larga Escala , Imunidade Inata/genética , Sirolimo/farmacologia
3.
Cell Signal ; 35: 84-94, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28336235

RESUMO

The small molecular weight GTPase Rab7 is a key regulator for late endosomal/lysosomal membrane trafficking, it was known that Rab7 is phosphorylated, but the corresponding kinase and the functional regulation of Rab7 phosphorylation remain unclear. We provide evidence here that Rab7 is a substrate of Src kinase, and is tyrosine-phosphorylated by Src, withY183 residue of Rab7 being the optimal phosphorylation site for Src. Further investigations demonstrated that the tyrosine phosphorylation of Rab7 depends on the guanine nucleotide binding activity of Rab7 and the activity of Src kinase. The tyrosine phosphorylation of Rab7 is physiologically induced by EGF, and impairs the interaction of Rab7 with RILP, consequently inhibiting EGFR degradation and sustaining Akt signaling. These results suggest that the tyrosine phosphorylation of Rab7 may be involved in coordinating membrane trafficking and cell signaling.


Assuntos
Transporte Proteico/genética , Transdução de Sinais/genética , Proteínas rab de Ligação ao GTP/genética , Quinases da Família src/genética , Proteínas Adaptadoras de Transdução de Sinal/genética , Membrana Celular/genética , Membrana Celular/metabolismo , Endossomos/genética , Fator de Crescimento Epidérmico/genética , Receptores ErbB/genética , Células HeLa , Humanos , Lisossomos/metabolismo , Fosforilação , Ligação Proteica , Proteólise , Tirosina/metabolismo
5.
Sci Rep ; 4: 7282, 2014 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-25445562

RESUMO

The HOPS complex serves as a tethering complex with GEF activity for Ypt7p in yeast to regulate late endosomal membrane maturation. While the role of HOPS complex is well established in yeast cells, its functional and mechanistic aspects in mammalian cells are less well defined. In this study, we report that RILP, a downstream effector of Rab7, interacts with HOPS complex and recruits HOPS subunits to the late endosomal compartment. Structurally, the amino-terminal portion of RILP interacts with HOPS complex. Unexpectedly, this interaction is independent of Rab7. VPS41 subunit of HOPS complex was defined to be the major partner for interacting with RILP. The carboxyl-terminal region of VPS41 was mapped to be responsible for the interaction. Functionally, either depletion of VPS41 by shRNA or overexpression of VPS41 C-terminal half retarded EGF-induced degradation of EGFR. These results suggest that interaction of RILP with HOPS complex via VPS41 plays a role in endocytic trafficking of EGFR.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas Nucleares/metabolismo , Transporte Proteico/fisiologia , Vesículas Transportadoras/metabolismo , Vesículas Transportadoras/fisiologia , Proteínas de Transporte Vesicular/metabolismo , Linhagem Celular Tumoral , Fator de Crescimento Epidérmico/metabolismo , Receptores ErbB , Células HeLa , Humanos , Células MCF-7
6.
PLoS One ; 9(5): e96242, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24788816

RESUMO

Rab29 (also referred as Rab7L1) is a novel Rab protein, and is recently demonstrated to regulate phagocytosis and traffic from the Golgi to the lysosome. However, its roles in membrane trafficking have not been investigated extensively. Our results in this study revealed that Rab29 is associated with the trans-Golgi network (TGN), and is essential for maintaining the integrity of the TGN, because inhibition of the activity of Rab29 or depletion of Rab29 resulted in fragmentation of the TGN marked by TGN46. Expression of the dominant negative form Rab29T21N or shRNA-Rab29 also altered the distribution of mannose-6-phosphate receptor (M6PR), and interrupted the retrograde trafficking of M6PR through monitoring the endocytosis of CD8-tagged calcium dependent M6PR (cdM6PR) or calcium independent M6PR (ciM6PR), but without significant effects on the anterograde trafficking of vesicular stomatitis virus G protein (VSV-G). Our results suggest that Rab29 is essential for the integrity of the TGN and participates in the retrograde trafficking of M6PRs.


Assuntos
Endossomos/metabolismo , Receptor IGF Tipo 2/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Proteínas rab1 de Ligação ao GTP/metabolismo , Rede trans-Golgi/metabolismo , Sequência de Aminoácidos , Western Blotting , Linhagem Celular , Retículo Endoplasmático/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Células HeLa , Humanos , Células MCF-7 , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Microscopia de Fluorescência , Dados de Sequência Molecular , Mutação , Transporte Proteico , Interferência de RNA , Receptor IGF Tipo 2/genética , Homologia de Sequência de Aminoácidos , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/metabolismo , Proteínas rab de Ligação ao GTP/genética , Proteínas rab1 de Ligação ao GTP/genética
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