Subcellular localization of bacteriophage PRD1 proteins in Escherichia coli

Bacteria possess an intricate internal organization resembling that of the eukaryotes. The complexity is especially prominent at the bacterial cell poles, which are also known to be the preferable sites for some bacteriophages to infect. Bacteriophage PRD1 is a well-known model serving as an ideal system to study structures and functions of icosahedral internal membrane-containing viruses. Our aim was to analyze the localization and interactions of individual PRD1 proteins in its native host Escherichia coli. This was accomplished by constructing a vector library for production of fluorescent fusion proteins. Analysis of solubility and multimericity of the fusion proteins, as well as their …

Sphingomyelin induces structural alteration in canine parvovirus capsid.

One of the essential steps in canine parvovirus (CPV) infection, the release from endosomal vesicles, is dominated by interactions between the virus capsid and the endosomal membranes. In this study, the effect of sphingomyelin and phosphatidyl serine on canine parvovirus capsid and on the phospholipase A(2) (PLA(2)) activity of CPV VP1 unique N-terminus was analyzed. Accordingly, a significant (P< or =0.05) shift of tryptophan fluorescence emission peak was detected at pH 5.5 in the presence of sphingomyelin, whereas at pH 7.4 a similar but minor shift was observed. This effect may relate to the exposure of VP1 N-terminus in acidic pH as well as to interactions between sphingomyelin and CP…

Sphingomyelin induces structural alteration in canine parvovirus capsid

One of the essential steps in canine parvovirus (CPV) infection, the release from endosomal vesicles, is dominated by interactions between the virus capsid and the endosomal membranes. In this study, the effect of sphingomyelin and phosphatidyl serine on canine parvovirus capsid and on the phospholipase A2 (PLA2) activity of CPV VP1 unique N-terminus was analyzed. Accordingly, a significant (P ≤ 0.05) shift of tryptophan fluorescence emission peak was detected at pH 5.5 in the presence of sphingomyelin, whereas at pH 7.4 a similar but minor shift was observed. This effect may relate to the exposure of VP1 N-terminus in acidic pH as well as to interactions between sphingomyelin and CPV. When…

Non-structural proteins P17 and P33 are involved in the assembly of the internal membrane-containing virus PRD1.

AbstractBacteriophage PRD1, which has been studied intensively at the structural and functional levels, still has some gene products with unknown functions and certain aspects of the PRD1 assembly process have remained unsolved. In this study, we demonstrate that the phage-encoded non-structural proteins P17 and P33, either individually or together, complement the defect in a temperature-sensitive GroES mutant of Escherichia coli for host growth and PRD1 propagation. Confocal microscopy of fluorescent fusion proteins revealed co-localisation between P33 and P17 as well as between P33 and the host chaperonin GroEL. A fluorescence recovery after photobleaching assay demonstrated that the diff…

Koiran parvoviruksen fysikaaliset ominaisuudet

Subcellular localization of bacteriophage PRD1 proteins in Escherichia coli

Bacteria possess an intricate internal organization resembling that of the eukaryotes. The complexity is especially prominent at the bacterial cell poles, which are also known to be the preferable sites for some bacteriophages to infect. Bacteriophage PRD1 is a well-known model serving as an ideal system to study structures and functions of icosahedral internal membrane-containing viruses. Our aim was to analyze the localization and interactions of individual PRD1 proteins in its native host Escherichia coli. This was accomplished by constructing a vector library for production of fluorescent fusion proteins. Analysis of solubility and multimericity of the fusion proteins, as well as their …

Interactions of virus proteins within the host cell

Kohtaamisia : yhteistoiminnan rakentuminen pariterapian reflektiivisten prosessien muutoskohdissa

Pro gradu -tutkielmassa tarkastelimme vuorovaikutustutkimuksen keinoin, miten yhteistoiminta rakentui hetki hetkeltä pariterapian reflektiivisten prosessien muutoskohdissa. Käyttämämme aineisto koostui Jyväskylän yliopiston Relationaalinen mieli -tutkimushankkeen yhteydessä kerätyistä videoidusta pariterapiaistunnoista. Kävimme läpi 62 videoitua pariterapiaistuntoa ja valikoimme niistä yhdeksän reflektiivisten prosessien aikaista muutoskohtaa. Muutoskohtina käsittelimme sellaisia reflektiivisen prosessin aikana esiintyneitä vuorovaikutusepisodeja, joihin sisältyi näkyviä vuorovaikutusresponsseja kummaltakin asiakaspariskunnan osapuolelta. Kehitimme monitoimijaisten vuorovaikutusepisodien li…