Ran GTPase Regulates Hemocytic Phagocytosis of Shrimp by Interaction with Myosin

Liu, Weifeng; Han, Fang; Zhang, Xiaobo

doi:10.1021/pr800840x

Cited by 97 publications

(82 citation statements)

References 35 publications

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“…To ensure that the shrimp were WSSV-free before the experiments, three shrimp were randomly selected for WSSV detection by use of WSSVspecific primers (5=-TTGGTTTCATGCCCGAGATT-3= and 5=-CCTTG GTCAGCCCCTTGA-3=). The virus-free shrimp were challenged with WSSV as described previously (16). Briefly, the tissues of WSSV-infected shrimp were homogenized in TN buffer (20 mM Tris-HCl, 400 mM NaCl, pH 7.4) at 0.1 g/ml.…”

Section: Methodsmentioning

confidence: 99%

“…To analyze the expression of the shrimp caspase 8 gene, quantitative real-time PCR was conducted as described previously (16). Shrimp ␤-actin was used as a control.…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

“…The sequence of the TaqMan probe was 5=-FAM-CTGGGTTACGAGTCTAA-TAMRA-3=. A linearized plasmid containing a 1,400-bp DNA fragment from the WSSV genome was quantified and serially diluted 10-fold as an internal standard for real-time PCR (16). The real-time PCR mixture (25 l) contained 12.5 l Ex Taq premix (TaKaRa, Japan), 1 l of the extracted DNA template or the internal standard plasmid, 0.5 l of 10 M (each) primers, and 0.5 l of 10 M TaqMan fluorogenic probe.…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

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Involvement of Viral MicroRNA in the Regulation of Antiviral Apoptosis in Shrimp

Huang

Cui

Zhang

2014

J Virol

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Viruses, in particular DNA viruses, generate microRNAs (miRNAs) to control the expression of host and viral genes. Due to their essential roles in virus-host interactions, viral miRNAs have attracted extensive investigations in recent years. To date, however, most studies on viral miRNAs have been conducted in cell lines. In this study, the viral miRNAs from white spot syndrome virus (WSSV) were characterized in shrimp in vivo. On the basis of our previous study and small RNA sequencing in this study, a total of 89 putative WSSV miRNAs were identified. As revealed by miRNA microarray analysis and Northern blotting, the expression of viral miRNAs was tissue specific in vivo. The results indicated that the viral miRNA WSSV-miR-N24 could target the shrimp caspase 8 gene, and this miRNA further repressed the apoptosis of shrimp hemocytes in vivo. As a result, the number of WSSV copies in shrimp in vivo was significantly increased compared with the control level (WSSV only). Therefore, our study presents the first report on the in vivo molecular events of viral miRNA in antiviral apoptosis.

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Section: Methodsmentioning

confidence: 99%

“…To analyze the expression of the shrimp caspase 8 gene, quantitative real-time PCR was conducted as described previously (16). Shrimp ␤-actin was used as a control.…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

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Involvement of Viral MicroRNA in the Regulation of Antiviral Apoptosis in Shrimp

Huang

Cui

Zhang

2014

J Virol

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“…These viruses employ Cdc42-hijacking tactics in their target cell entry processes (14). Some GTPases show antiviral functions by interacting with actin in shrimp; for example, the GTPase Rab6 interacts with actin to regulate phagocytosis in Marsupenaeus japonicus hemocytes (15), and the GTPase Ran is involved in the hemocytic phagocytosis of white spot syndrome virus (WSSV) through its interaction with myosin (16). Rac1, the other member of the Rho family, plays a role in the antiviral and antibacterial innate immunity of shrimp (17).…”

mentioning

confidence: 99%

Interaction of the Small GTPase Cdc42 with Arginine Kinase Restricts White Spot Syndrome Virus in Shrimp

Jiang

Wei

et al. 2017

J Virol

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Many types of small GTPases are widely expressed in eukaryotes and have different functions. As a crucial member of the Rho GTPase family, Cdc42 serves a number of functions, such as regulating cell growth, migration, and cell movement. Several RNA viruses employ Cdc42-hijacking tactics in their target cell entry processes. However, the function of Cdc42 in shrimp antiviral immunity is not clear. In this study, we identified a Cdc42 protein in the kuruma shrimp (Marsupenaeus japonicus) and named it MjCdc42. MjCdc42 was upregulated in shrimp challenged by white spot syndrome virus (WSSV). The knockdown of MjCdc42 and injection of Cdc42 inhibitors increased the proliferation of WSSV. Further experiments determined that MjCdc42 interacted with an arginine kinase (MjAK). By analyzing the binding activity and enzyme activity of MjAK and its mutant, ΔMjAK, we found that MjAK could enhance the replication of WSSV in shrimp. MjAK interacted with the envelope protein VP26 of WSSV. An inhibitor of AK activity, quercetin, could impair the function of MjAK in WSSV replication. Further study demonstrated that the binding of MjCdc42 and MjAK depends on Cys 271 of MjAK and suppresses the WSSV replication-promoting effect of MjAK. By interacting with the active site of MjAK and suppressing its enzyme activity, MjCdc42 inhibits WSSV replication in shrimp. Our results demonstrate a new function of Cdc42 in the cellular defense against viral infection in addition to the regulation of actin and phagocytosis, which has been reported in previous studies. IMPORTANCEThe interaction of Cdc42 with arginine kinase plays a crucial role in the host defense against WSSV infection. This study identifies a new mechanism of Cdc42 in innate immunity and enriches the knowledge of the antiviral innate immunity of invertebrates.

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“…Some documents indicated that small GTPases are related to virus infection in the shrimp (Wu et al 2007;Liu et al 2009;Zhang et al 2010). Also, the small GTPases of zebra fish have provided a firm basis of an innate immune system in vertebrates (Salas-Vidal et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Cloning and characterization of ADP-ribosylation factor 1b from the olive flounder Paralichthys olivaceus

Son

Jang

et al. 2017

Fish Aquatic Sci

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Small GTPases are well known as one of the signal transduction factors of immune systems. The ADP-ribosylation factors (ARFs) can be classified into three groups based on the peptide sequence, protein molecular weight, gene structure, and phylogenetic analysis. ARF1 recruits coat proteins to the Golgi membranes when it is bound to GTP. The class I duplicated ARF gene was cloned and characterized from the olive flounder (Paralichthys olivaceus) for this study. PoARF1b contains the GTP-binding motif and the switch 1 and 2 regions. PoARF1b and PoARF1b mutants were transfected into a Hirame natural embryo cell to determine the distribution of its GDP/GTP-bound state; consequently, it was confirmed that PoARF1b associates with the Golgi body when it is in a GTP-binding form. The results of the qPCR-described PoARF1b were expressed for all of the P. olivaceus tissues. The authors plan to study the gene expression patterns of PoARF1b in terms of immunity challenges.

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Ran GTPase Regulates Hemocytic Phagocytosis of Shrimp by Interaction with Myosin

Cited by 97 publications

References 35 publications

Involvement of Viral MicroRNA in the Regulation of Antiviral Apoptosis in Shrimp

Involvement of Viral MicroRNA in the Regulation of Antiviral Apoptosis in Shrimp

Interaction of the Small GTPase Cdc42 with Arginine Kinase Restricts White Spot Syndrome Virus in Shrimp

Cloning and characterization of ADP-ribosylation factor 1b from the olive flounder Paralichthys olivaceus

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