Hostname: page-component-78c5997874-s2hrs Total loading time: 0 Render date: 2024-11-10T10:42:22.406Z Has data issue: false hasContentIssue false
  • English
  • Français

Structural modelling studies and immunoprophylactic potential of Brugia malayi DEAD Box RNA helicase

Published online by Cambridge University Press:  07 June 2013

MEGHNA SINGH ,
NIDHI SHRIVASTAVA ,
UZMA SAQIB ,
MOHAMMAD IMRAN SIDDIQI  and
SHAILJA MISRA-BHATTACHARYA
MEGHNA SINGH
Affiliation:
Division of Parasitology, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram extension, Sitapur Road, Lucknow 226021, Uttar Pradesh, India
NIDHI SHRIVASTAVA
Affiliation:
Division of Parasitology, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram extension, Sitapur Road, Lucknow 226021, Uttar Pradesh, India
UZMA SAQIB
Affiliation:
Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram extension, Sitapur Road, Lucknow 226021, Uttar Pradesh, India
MOHAMMAD IMRAN SIDDIQI
Affiliation:
Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram extension, Sitapur Road, Lucknow 226021, Uttar Pradesh, India
SHAILJA MISRA-BHATTACHARYA*
Affiliation:
Division of Parasitology, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram extension, Sitapur Road, Lucknow 226021, Uttar Pradesh, India
*
*Corresponding author: Division of Parasitology, Central Drug Research Institute, Lucknow-226021, UP, India. Tel: +91 522 2612411 18. PABX 4221/4224. Fax: +91 522 2623405/2623938/2629504. E-mail: shailja_cdri@rediffmail.com, shailjacdri@yahoo.com
Get access Rights & Permissions [Opens in a new window]

Summary

DEAD Box RNA helicases are essential enzymes that are involved in RNA metabolic processes such as transcription, pre-mRNA splicing, translation initiation and RNA decay. We have previously over-expressed and biochemically characterized an immunodominant cDNA clone encoding DEAD box RNA helicase (BmL3-Helicase) isolated by immunoscreening of the larval stage cDNA library of Brugia malayi. In the current study, the 3D structure was determined and the immunoprophylactic efficacy of BmL3-Helicase was investigated by immunizing Mastomys coucha with the recombinant protein and subsequently challenging with B. malayi infective larvae. The immunization had an adverse outcome on the establishment of challenged larvae resulting in a 67·4% reduction in adult parasite recovery, a 86·7% decrease in the microfilarial density and profound sterility of the recovered female worms. The immune response thus generated was investigated by measuring the levels of specific antibodies including IgG subclasses, reactive oxygen species and cytokines.

Keywords

BmL3-Helicasehomology modellingimmunoprophylaxis
Type
Research Article
Information
Parasitology , Volume 140 , Issue 8 , July 2013 , pp. 1016 - 1025
Copyright
Copyright © Cambridge University Press 2013 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Altschul, S. F., Thomas, L. M., Alejandro, A. S., Jinghui, Z., Zheng, Z., Webb, M. and David, J. L. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research 25, 3389–3402.CrossRefGoogle ScholarPubMed
Chandrashekar, R., Rao, U. R., Parab, Ρ. Β. and Subrahmanyam, D. (1985). Brugia malayi: serum-dependent cell-mediated reactions to microfilariae. Southeast Asian Journal of Tropical Medicine and Public Health 16, 15–21.Google ScholarPubMed
Devaney, E. and Osborne, J. (2000). The third-stage larva (L3) of Brugia: its role in immune modulation and protective immunity. Microbes and Infection 2, 1363–1371.CrossRefGoogle ScholarPubMed
Ghedin, E., Wang, S., Spiro, D., Caler, E., Zhao, Q., Crabtree, J., Allen, J. E., Delcher, A. L., Guiliano, D. B., Miranda-Saavedra, D., Angiuoli, S. V., Creasy, T. et al. (2007). Draft genome of the filarial nematode parasite Brugia malayi. Science 317, 1756–1760.CrossRefGoogle ScholarPubMed
Gregory, W. F., Atmadja, A. K., Allen, J. E. and Maizels, R. M. (2000). The abundant larval transcript-1 and -2 genes of Brugia malayi encode stage-specific candidate vaccine antigens for filariasis. Infection and Immunity 68, 4174–4179.CrossRefGoogle ScholarPubMed
Kolaskar, A. S. and Tongaonkar, P. C. (1990). A semi-empirical method for prediction of antigenic determinants on protein antigens. FEBS Letters 276, 172–174.CrossRefGoogle ScholarPubMed
Laskowski, R. A., MacArthur, M. W., Moss, D. S. and Thornton, J. M. (1993). PROCHECK: a program to check the stereochemical quality of protein structures. Journal of Applied Crystallography 26, 283–291.CrossRefGoogle Scholar
Pause, A., Méthot, N. and Sonenberg, N. (1993). The HRIGRXXR region of the DEAD box RNA helicase eukaryotic translation factor 4A is required for RNA binding and ATP hydrolysis. Molecular and Cellular Biology 13, 6789–6798.Google ScholarPubMed
Rajan, T. V., Porte, P., Yates, J. A., Keefer, L. and Shultz, L. D. (1996). Role of nitric oxide in host defense against an extracellular, metazoan parasite, Brugia malayi. Infection and Immunity 64, 3351–3353.CrossRefGoogle ScholarPubMed
Sali, A. and Blundell, T. L. (1993). Comparative protein modeling by satisfaction of spatial restraints. Journal of Molecular Biology 234, 779–815.CrossRefGoogle ScholarPubMed
Singh, H. and Raghava, G. P. S. (2001). ProPred: prediction of HLA-DR binding sites. Bioinformatics 17, 1236–1237.CrossRefGoogle ScholarPubMed
Singh, M., Shakya, S., Soni, V. K., Dangi, A., Kumar, N. and Bhattacharya, S. M. (2009 a). The n-Hexane and chloroform fractions of Piper betle L. trigger different arms of immune responses in BALB/c mice. International Immunopharmacology 9, 716–728.CrossRefGoogle ScholarPubMed
Singh, M., Srivastava, K. K. and Bhattacharya, S. M. (2009 b). Molecular cloning and characterization of a novel immunoreactive ATPase/RNA helicase in human filarial parasite Brugia malayi. Parasitological Research 104, 753–761.CrossRefGoogle ScholarPubMed
Singh, U., Misra, S., Muthy, P. K., Katiyar, J. C., Agrawal, A. and Sircar, A. R. (1997). Immunoreactive molecules of Brugia malayi and their diagnostic potential. Serodiagnosis and Immunotherapy in Infectious Disease 8, 207–212.CrossRefGoogle Scholar
Taylor, M. J., Hoerauf, A. and Bockarie, M. (2010). Lymphatic filariasis and onchocerciasis. Lancet 376, 1175–1185.CrossRefGoogle ScholarPubMed
Thomas, G. R., Crossan, M. C. and Selkirk, M. E. (1997). Cytostatic and cytotoxic effects of activated macrophages and nitric oxide donors on Brugia malayi. Infection and Immunity 65, 2732–2739.CrossRefGoogle ScholarPubMed
Vedi, S., Dangi, A., Hajela, K. and Misra-Bhattacharya, S. (2008). Vaccination with 73 kDa recombinant heavy chain myosin generates high level of protection against Brugia malayi challenge in jird and mastomys models. Vaccine 26, 5997–6005.CrossRefGoogle Scholar