Virus-like particle nanoreactors: programmed en capsulation of the thermostable CelB glycosidase inside the P22 capsid
Patterson, D.P. ; Schwarz, B. ; El-Boubbou, K. ; Oost, J. van der; Prevelige, P.E. ; Douglas, T. - \ 2012
Soft Matter 8 (2012)39. - ISSN 1744-683X - p. 10158 - 10166.
beta-glucosidase celb - archaeon pyrococcus-furiosus - protein cages - scaffolding protein - bacteriophage p22 - silica immobilization - enzyme immobilization - gold nanoparticles - functional domains - escherichia-coli
Self-assembling biological systems hold great potential for the synthetic construction of new active soft nanomaterials. Here we demonstrate the hierarchical bottom-up assembly of bacteriophage P22 virus-like particles (VLPs) that encapsulate the thermostable CelB glycosidase creating catalytically active nanoreactors. The in vivo assembly and encapsulation produces P22 VLPs with a high packaging density of the tetrameric CelB, but without loss of enzyme activity or the ability of the P22 VLP to undergo unique morphological transitions that modify the VLPs internal volume and shell porosity. The P22 VLPs encapsulating CelB are also shown to retain a high percentage of the enzyme activity upon being embedded and immobilized in a polymeric matrix
Transcriptome analysis of infection of the archaeon Sulfolobus solfataricus with Sulfolobus turreted icosahedral virus
Ortmann, A.C. ; Walther, J. ; Brumfield, S.K. ; McInnerney, K. ; Brouns, S.J.J. ; Werken, H.J.G. van de; Bothner, B. ; Douglas, T. ; Oost, J. van der; Young, M.J. - \ 2008
Journal of Virology 82 (2008)10. - ISSN 0022-538X - p. 4874 - 4883.
gene-expression - electron microscopy - particle ssv1 - dna - protein - genome - host - replication - microarrays - chromosome
Microarray analysis of infection by Sulfolobus turreted icosahedral virus (STIV) revealed insights into the timing and extent of virus transcription, as well as differential regulation of host genes. Using a microarray containing genes from both the host and the virus, the infection cycle of STIV was studied. Following infection of Sulfolobus solfataricus strain 2-2-12 with STIV, transcription of virus genes was first detected at 8 h postinfection (p.i.), with a peak at 24 h p.i. Lysis of cells was first detected at 32 h p.i. There was little temporal control of the transcription of virus genes, although the three open reading frames on the noncoding strand were transcribed later in the infection process. During the infection, 177 host genes were determined to be differentially expressed, with 124 genes up-regulated and 53 genes down-regulated. The up-regulated genes were dominated by genes associated with DNA replication and repair and those of unknown function, while the down-regulated genes, mostly detected at 32 h p.i., were associated with energy production and metabolism. Examination of infected cells by transmission electron microscopy revealed alterations in cell ultrastructure consistent with the microarray analysis. The observed patterns of transcription suggest that up-regulated genes are likely used by the virus to reprogram the cell for virus replication, while the down-regulated genes reflect the imminent lysis of the cells.