scholarly journals A Novel Digestive Proteinase Lipase Member H-A in Bombyx mori Contributes to Digestive Juice Antiviral Activity against B. mori Nucleopolyhedrovirus

Insects ◽  
2020 ◽  
Vol 11 (3) ◽  
pp. 154 ◽  
Author(s):  
Shang-Zhi Zhang ◽  
Lin-Bao Zhu ◽  
Ling-Ling You ◽  
Jie Wang ◽  
Hui-Hua Cao ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Bombyx Mori ◽  
Treated Group ◽  
Middle Part ◽  
Expression Level ◽  
Label Free ◽  
Proteomics Data ◽  
Gene Encoding ◽  
Digestive Juice ◽  
Bmn Cells

Previous studies have revealed that some proteins in Bombyx mori larvae digestive juice show antiviral activity. Here, based on the label-free proteomics data, BmLipase member H-A (BmLHA) was identified as being involved in the response to BmNPV infection in B. mori larvae digestive juice. In the present study, a gene encoding the BmLHA protein in B. mori was characterized. The protein has an open reading fragment of 999 bp, encoding a predicted 332 amino acid residue-protein with a molecular weight of approximately 35.9 kDa. The phylogenetic analysis revealed that BmLHA shares a close genetic distance with Papilio xuthus Lipase member H-A. BmLHA was highly expressed in the middle part of the B. mori gut, and the expression level increased with instar rising in larvae. There was higher expression of BmLHA in A35 than in P50 strains, and it was upregulated in both A35 and P50 strains, following BmNPV infection. The expression level of VP39 decreased significantly in appropriate recombinant-BmLHA-treated groups compared with the PBS-treated group in B. mori larvae and BmN cells. Meanwhile, overexpression of BmLHA significantly reduced the infectivity of BmNPV in BmN cells. These results indicated that BmLHA did not have digestive function but had anti-BmNPV activity. Taken together, our work provides valuable data for the clarification of the molecular characterization BmLHA and supplements research on proteins of anti-BmNPV activity in B. mori.

IN VITRO ANTIVIRAL ACTIVITY OF AN ALKALINE TRYPSIN FROM THE DIGESTIVE JUICE OF Bombyx mori LARVAE AGAINST NUCLEOPOLYHEDROVIRUS

2012 ◽  
Vol 81 (2) ◽  
pp. 90-104 ◽  
Author(s):  
Kangayam M. Ponnuvel ◽  
Krishnappa Nithya ◽  
Sasibhushan Sirigineedi ◽  
Arvind K. Awasthi ◽  
Minoru Yamakawa
Keyword(s):  
Antiviral Activity ◽  
Bombyx Mori ◽  
Digestive Juice

Decrease in the expression level of the gene encoding the putative Bombyx mori bidensovirus receptor during virus infection

2018 ◽  
Vol 163 (12) ◽  
pp. 3327-3338 ◽  
Author(s):  
Katsuhiko Ito ◽  
Takeshi Fujii ◽  
Takeshi Yokoyama ◽  
Keiko Kadono-Okuda
Keyword(s):  
Virus Infection ◽  
Bombyx Mori ◽  
Expression Level ◽  
Gene Encoding

scholarly journals Antiviral activity of a serine protease from the digestive juice of Bombyx mori larvae against nucleopolyhedrovirus

Virology ◽  
2004 ◽  
Vol 321 (1) ◽  
pp. 154-162 ◽  
Author(s):  
Hiroshi Nakazawa ◽  
Eiko Tsuneishi ◽  
Kangayam M Ponnuvel ◽  
Seiichi Furukawa ◽  
Ai Asaoka ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Bombyx Mori ◽  
Serine Protease ◽  
Digestive Juice

scholarly journals A Lipase Isolated from the Silkworm Bombyx mori Shows Antiviral Activity against Nucleopolyhedrovirus

2003 ◽  
Vol 77 (19) ◽  
pp. 10725-10729 ◽  
Author(s):  
Kangayam M. Ponnuvel ◽  
Hiroshi Nakazawa ◽  
Seiichi Furukawa ◽  
Ai Asaoka ◽  
Jun Ishibashi ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Antiviral Activity ◽  
Bombyx Mori ◽  
Northern Blot Analysis ◽  
Digestive Enzyme ◽  
Homology Search ◽  
Digestive Juice ◽  
Strong Antiviral Activity ◽  
Silkworm Bombyx Mori

ABSTRACT A protein showing strong antiviral activity against Bombyx mori nucleopolyhedrovirus (BmNPV) was purified from the digestive juice of B. mori larvae. A homology search of the deduced amino acid sequence of the protein cDNA revealed 56% homology with Drosophila melanogaster lipase and 21% homology with human lipase. As lipase activity of the protein was confirmed in vitro, this protein was designated Bmlipase-1. Northern blot analysis showed that the Bmlipase-1 gene is expressed in the midgut but not in other tissues, nor is it activated by BmNPV infection. In addition, the Bmlipase-1 gene was shown not to be expressed in the molting and wandering stages, indicating that the gene is hormonally regulated. Our results suggest that an insect digestive enzyme has potential as a physiological barrier against BmNPV at the initial site of viral infection.

scholarly journals Proteomic Analysis of Capacitated and Non-Capacitated Spermatozoa of Yanbian Yellow Cattle

2020 ◽  
Author(s):  
Xuan Chen ◽  
Zuochen Li ◽  
Yanqiu Lv ◽  
Yichao Xu ◽  
Mimi Cheng ◽  
...  
Keyword(s):  
Protein Transport ◽  
Expression Level ◽  
Molecular Processes ◽  
Proteomics Data ◽  
Pathway Enrichment ◽  
Protein Protein Interaction ◽  
Go Enrichment ◽  
Yellow Cattle ◽  
Specific Proteins ◽  
The Difference

Abstract Background: Sperm capacitation is a process which occurs prior to fertilization, and is essential for producing high-quality living embryos. The main purpose of this study was to explore the difference of proteomics between capacitated and non-capacitated sperm of Yanbian yellow cattle. Bioinformatic analyses of LC-MS/MS data included GO enrichment, KEGG pathway enrichment, and protein-protein interaction (PPI) analysis. Results: The results revealed 23 specific proteins in the capacitated group and 345 in the non-capacitated group. Compared with non-capacitated sperm, capacitated sperm exhibited 89 upregulated proteins and 509 downregulated proteins. Western blotting was used to confirm our proteomics data. The expression level of PSMD1 in the capacitated sperm group was significantly lower than that in the non-capacitated sperm group, and the expression level of HSPA5 was significantly higher than in the non-capacitated sperm group. Conclusions: Our results revealed that many proteins were differentially expressed between capacitated and non-capacitated sperm, particularly those involved in the proteasome signaling and protein transport signaling pathways. This work enhances our understanding of molecular processes involved in sperm viability in Yanbian yellow cattle, and provides a framework for future studies.

scholarly journals Bombyx mori β-1,3-Glucan Recognition Protein 4 (BmβGRP4) Could Inhibit the Proliferation of B. mori Nucleopolyhedrovirus through Promoting Apoptosis

Insects ◽  
2021 ◽  
Vol 12 (8) ◽  
pp. 743
Author(s):  
Jie Wang ◽  
Lin-Bao Zhu ◽  
Yan Ma ◽  
Ying-Xue Liu ◽  
Hui-Hua Cao ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Protein Gene ◽  
Phosphatase And Tensin Homolog ◽  
Reading Frame ◽  
Larval Stages ◽  
Tensin Homolog ◽  
Transcriptome Database ◽  
Recognition Protein ◽  
Apoptosis Gene ◽  
Bmn Cells

β-1,3-glucan recognition proteins (βGRPs) as pattern recognition receptors (PRRs) play an important role in recognizing various pathogens and trigger complicated signaling pathways in insects. In this study, we identified a Bombyx mori β-1,3-glucan recognition protein gene named BmβGRP4, which showed differential expression, from a previous transcriptome database. The full-length cDNA sequence was 1244 bp, containing an open reading frame (ORF) of 1128 bp encoding 375 amino acids. BmβGRP4 was strongly expressed in the larval stages and highly expressed in the midgut of B. mori larvae in particular. After BmNPV infection, the expression of BmβGRP4 was reduced significantly in the midgut. Furthermore, a significant increase in the copy number of BmNPV was observed after the knockdown of BmβGRP4 in 5th instar larvae, while the overexpression of BmβGRP4 suppressed the proliferation of BmNPV in BmN cells. Subsequently, the expression analysis of several apoptosis-related genes and observation of the apoptosis morphology demonstrated that overexpression of BmβGRP4 facilitated apoptosis induced by BmNPV in BmN cells. Moreover, BmβGRP4 positively regulated the phosphatase and tensin homolog gene (BmPTEN), while expression of the inhibitor of apoptosis gene (BmIAP) was negatively regulated by BmβGRP4. Hence, we hypothesize that BmNPV infection might suppress BmPTEN and facilitate BmIAP to inhibit cell apoptosis by downregulating the expression of BmβGRP4 to escape host antiviral defense. Taken together, these results show that BmβGRP4 may play a role in B. mori response to BmNPV infection and lay a foundation for studying its functions.

scholarly journals Identification of an enhancer-like element in the polyhedrin gene upstream region of Bombyx mori nucleopolyhedrovirus

2001 ◽  
Vol 82 (11) ◽  
pp. 2811-2819 ◽  
Author(s):  
Asha Acharya ◽  
Karumathil P. Gopinathan
Keyword(s):  
Bombyx Mori ◽  
Reporter Gene ◽  
Fold Increase ◽  
Firefly Luciferase ◽  
Upstream Region ◽  
Upstream Sequence ◽  
Gene Encoding ◽  
Viral Promoters ◽  
Enhancer Sequences ◽  
In Cis

A series of deletions in the upstream region of the gene encoding polyhedrin (polh) of Bombyx mori nucleopolyhedrovirus (BmNPV) were generated in plasmid constructs and tested for transcription. In transient transfection assays in Bombyx mori-derived BmN cells with firefly luciferase as the reporter gene, a 293 bp fragment located 1·0 kb upstream with respect to the +1 ATG of polh showed 10-fold enhancement in expression from the minimal promoter. This increase in reporter activity was observed only when the fragment was positioned in cis with respect to the promoter and not in trans. The stimulation of reporter gene expression was independent of the orientation of the fragment and was due to increased transcription from the promoter. When placed upstream of another promoter, the viral very late gene p10 promoter, the enhancer brought about a 2-fold increase in expression. The region encompassing the enhancer was itself transcriptionally active, and transcripts corresponding to both of the encoded ORFs (N-terminal regions of ORF453 and ORF327, located in opposite orientations) were detected. Two AP1 sites (TGACTCG) in the 293 bp fragment did not appear to contribute to the enhancer function. Since repeat motifs, the hallmark of conventional enhancer sequences, were absent from this fragment, it is designated as an enhancer-like element. The influence of this region of the polh upstream sequence on expression from strong, very late viral promoters has not been reported previously.

Characterization of a gene encoding prohibitin in silkworm, Bombyx mori

Gene ◽  
2012 ◽  
Vol 502 (2) ◽  
pp. 118-124 ◽  
Author(s):  
Zhengbing Lv ◽  
Xufang Zhang ◽  
Lili Liu ◽  
Jian Chen ◽  
Zuoming Nie ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Gene Encoding ◽  
Silkworm Bombyx Mori

The mycobacterial efflux pump EfpA can induce high drug tolerance to many anti-tuberculosis drugs, including moxifloxacin, in Mycobacterium smegmatis

2021 ◽  
Author(s):  
Deepika Rai ◽  
Sarika Mehra
Keyword(s):  
Mycobacterium Smegmatis ◽  
Efflux Pump ◽  
Drug Tolerance ◽  
Inducible Promoter ◽  
Major Facilitator Superfamily ◽  
Expression Level ◽  
Efflux Pump Inhibitor ◽  
Gene Encoding ◽  
Active Efflux ◽  
Major Facilitator

Active efflux of drugs across the membrane is a major survival strategy of bacteria against many drugs. In this work, we characterize an efflux pump EfpA, from the major facilitator superfamily, that is highly conserved among both slow growing and fast-growing mycobacterium species and has been found to be upregulated in many clinical isolates of Mycobacterium tuberculosis . The gene encoding EfpA from Mycobacterium smegmatis was over-expressed under both constitutive and an inducible promoter. Expression of efpA gene under both the promoters resulted in greater than 32-fold increased drug tolerance of M. smegmatis cells to many first-line (rifampicin, isoniazid and streptomycin) and second-line (amikacin) anti-tuberculosis drugs. Notably, drug tolerance of M. smegmatis cells to moxifloxacin increased by more than 180-fold when efpA was over-expressed. The increase in minimum inhibitory concentration (MIC) correlated with the decreased uptake of drugs including norfloxacin, moxifloxacin and ethidium bromide and the high MIC could be reversed in the presence of an efflux pump inhibitor. A correlation was observed between the MIC of drugs and the efflux pump expression level, suggesting that the latter could be modulated by varying the expression level of the efflux pump. The expression of high levels of efpA did not impact the fitness of the cells when supplemented with glucose.The efpA gene is conserved across both pathogenic and non-pathogenic mycobacteria. The efpA gene from the Mycobacterium bovis BCG/ M. tuberculosis , which is 80% identical to efpA from M. smegmatis , also led to decreased antimicrobial efficacy to many drugs, although the fold-change was lower. When over-expressed in M. bovis BCG, an 8-fold higher drug tolerance to moxifloxacin was observed . This is the first report of an efflux pump from mycobacterium species that leads to higher drug tolerance to moxifloxacin, a promising new drug for the treatment of tuberculosis.

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