Dataset Information

Intracellular localization of Crimean-Congo Hemorrhagic Fever (CCHF) virus glycoproteins.

ABSTRACT: BACKGROUND: Crimean-Congo Hemorrhagic Fever virus (CCHFV), a member of the genus Nairovirus, family Bunyaviridae, is a tick-borne pathogen causing severe disease in humans. To better understand the CCHFV life cycle and explore potential intervention strategies, we studied the biosynthesis and intracellular targeting of the glycoproteins, which are encoded by the M genome segment. RESULTS: Following determination of the complete genome sequence of the CCHFV reference strain IbAr10200, we generated expression plasmids for the individual expression of the glycoproteins GN and GC, using CMV- and chicken beta-actin-driven promoters. The cellular localization of recombinantly expressed CCHFV glycoproteins was compared to authentic glycoproteins expressed during virus infection using indirect immunofluorescence assays, subcellular fractionation/western blot assays and confocal microscopy. To further elucidate potential intracellular targeting/retention signals of the two glycoproteins, GFP-fusion proteins containing different parts of the CCHFV glycoprotein were analyzed for their intracellular targeting. The N-terminal glycoprotein GN localized to the Golgi complex, a process mediated by retention/targeting signal(s) in the cytoplasmic domain and ectodomain of this protein. In contrast, the C-terminal glycoprotein GC remained in the endoplasmic reticulum but could be rescued into the Golgi complex by co-expression of GN. CONCLUSION: The data are consistent with the intracellular targeting of most bunyavirus glycoproteins and support the general model for assembly and budding of bunyavirus particles in the Golgi compartment.

SUBMITTER: Haferkamp S

PROVIDER: S-EPMC1090624 | biostudies-literature | 2005

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Intracellular localization of Crimean-Congo Hemorrhagic Fever (CCHF) virus glycoproteins.

Haferkamp Sebastian S Fernando Lisa L Schwarz Tino F TF Feldmann Heinz H Flick Ramon R

Virology journal 20050425

<h4>Background</h4>Crimean-Congo Hemorrhagic Fever virus (CCHFV), a member of the genus Nairovirus, family Bunyaviridae, is a tick-borne pathogen causing severe disease in humans. To better understand the CCHFV life cycle and explore potential intervention strategies, we studied the biosynthesis and intracellular targeting of the glycoproteins, which are encoded by the M genome segment.<h4>Results</h4>Following determination of the complete genome sequence of the CCHFV reference strain IbAr10200 ...[more]

PMID: 15850490

Similar Datasets

Project description:Crimean-Congo hemorrhagic fever (CCHF) virus is the cause of an important tick-borne disease of humans throughout regions of Africa, Europe, and Asia. Like other members of the genus Nairovirus, family Bunyaviridae, the CCHF virus M genome RNA segment encodes the virus glycoproteins. Sequence analysis of the CCHF virus (Matin strain) M RNA segment revealed one major open reading frame that potentially encodes a precursor polyprotein 1,689 amino acids (aa) in length. Comparison of the deduced amino acid sequences of the M-encoded polyproteins of Nigerian, Pakistani, and Chinese CCHF virus strains revealed two distinct protein regions. The carboxyl-terminal 1,441 aa are relatively highly conserved (up to 8.4% identity difference), whereas the amino-terminal 243 to 248 aa are highly variable (up to 56.4% identity difference) and have mucin-like features, including a high serine, threonine, and proline content (up to 47.3%) and a potential for extensive O-glycosylation. Analysis of released virus revealed two major structural glycoproteins, G2 (37 kDa) and G1 (75 kDa). Virus protein analysis by various techniques, including pulse-chase analysis and/or reactivity with CCHF virus-specific polyclonal and antipeptide antibodies, demonstrated that the 140-kDa (which contains the mucin-like region) and 85-kDa nonstructural proteins are the precursors of the mature G2 and G1 proteins, respectively. The amino termini of the CCHF virus (Matin strain) G2 and G1 proteins were established by microsequencing to be equivalent to aa 525 and 1046, respectively, of the encoded polyprotein precursor. The tetrapeptides RRLL and RKPL are immediately upstream of the cleavage site for mature G2 and G1, respectively. These are completely conserved among the predicted polyprotein sequences of all the CCHF virus strains and closely resemble the tetrapeptides that represent the major cleavage recognition sites present in the glycoprotein precursors of arenaviruses, such as Lassa fever virus (RRLL) and Pichinde virus (RKLL). These results strongly suggest that CCHF viruses (and other members of the genus Nairovirus) likely utilize the subtilase SKI-1/S1P-like cellular proteases for the major glycoprotein precursor cleavage events, as has recently been demonstrated for the arenaviruses.

Project description:BackgroundCrimean Congo Hemorrhagic (CCHF) is a deadly tick born disease caused by a virus of genus Nairovirus and is endemic in the Middle East, Asia, Africa, and Eastern areas of Europe. Pakistan is a CCHF endemic country with a constant threat of sporadic outbreaks. Health care workers are more prone to CCHF, hence, it is a prerequisite for members of the healthcare team to stay abreast with current knowledge and display positive attitude and perception. This study assessed the medical and pharmacy students' preparedness level in terms of CCHF control and management.MethodsA total of 900 consenting students were selected randomly, who completed a predesigned and validated questionnaire which assessed the participant's general knowledge, emergency preparedness control and management of CCHF. Data were analyzed by SPSS (IBM SPSS version 21). For data analysis percentages, P-value, t-test, the independent sample mean, Whitney U test, Kruskal-Wallis test, Logistic regression, and Spearman correlation were utilized.ResultsAmong 900 study respondents, 68% were females and 32% were males, out of which physicians (MBBS) students were 48.4%, and pharmacists students were 51.6%. Majority of the respondents 39.9% were from age group of 22-25?years. Overall 43% healthcare students demonstrated good knowledge about disease causes, transmission, and treatment options. Additionally, 81% of the study participants showed positive attitude, whereas, 69% students demonstrated positive perceptions. The correlation coefficient showed positive correlation between attitude- perception (r =?0.268, p value?=?0.000), knowledge- attitude (r =?0.234, p value?=?0.000) and knowledge- perception (r =?0.257, p value?=?0.000).ConclusionsKnowledge gaps were observed which is alarming. These gaps were multifactorial and mainly due to lack of knowledge, poor motivation, and old syllabus which needs to be addressed. The study results show that it is crucial to evaluate current curriculum and also showing a dire need of awareness seminars, conferences workshops to highlight and educate about the current endemic disease to future health care professionals.

Dataset Information

Intracellular localization of Crimean-Congo Hemorrhagic Fever (CCHF) virus glycoproteins.

Publications

Intracellular localization of Crimean-Congo Hemorrhagic Fever (CCHF) virus glycoproteins.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure