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Herenyi, L.

Publications and source records attributed to Herenyi, L..

3 recordsLinked to original sources

Flexible Motion of T7 bacteriophage Tail Fibers Suggest a Dynamic Viral Infection Mechanism

Viruses are nanoscale infectious agents capable of specifically targeting and reprograming host cells. A unique group of viruses, bacteriophages, have regained popularity in research partly due to the rising number of multidrug-resistant bacterial infections. Phages could potentially replace antibiotics, but only if we understand every detail of their structure and infection cycle. T7 bacteriophages are a group of dsDNA viruses, which infect E. coli bacteria. T7 virions are comprised of an icosahedral protein shell which encapsulates the genomic DNA, and a tail-fiber complex which is primarily used for target recognition and DNA injection. The virus has six" L"-shaped, [~]40 nm long fibers (gp17 protein trimers) attached to the tail-tube, which are thought to be essential for initial host recognition and possibly surface exploration. Using high-speed atomic force microscopy (HS-AFM) and molecular dynamics (MD) simulations combined with small angle X-ray scattering (SAXS) we observed the molecular structure and movements of isolated tail fibers. Firstly, we have identified a hinge region within the fibers, which makes them highly flexible, allowing the bending of their distal region. Furthermore, we have observed the dynamic triple helical coiled coil structure of the proximal region, which would allow fiber rotation. These two points of flexibility allow a more efficient and highly dynamic host recognition and virus anchoring process. The observed flexibility might allow host surface exploration by walking. Such flexibility in the host recognition machinery may not be unique to T7 bacteriophages, getting us one step closer to understanding the intricate details of virus-host interactions.

biophysics↗

TMPyP binding evokes a complex, tunable nanomechanical response in DNA

TMPyP is a porphyrin capable of DNA binding and used in photodynamic therapy and G-quadruplex stabilization. Despite its broad applications, the effect of TMPyP on DNA nanomechanics is unknown. Here we investigated, by manipulating {lambda}-phage DNA with optical tweezers combined with microfluidics, how TMPyP influences DNA nanomechanics across a wide range of TMPyP concentration (5-5120 nM), mechanical force (0-100 pN), NaCl concentration (0.01-1 M) and pulling rate (0.2-20 m/s). Complex responses were recorded, for the analysis of which we introduced a simple mathematical model. TMPyP binding leads to the lengthening and softening of dsDNA. dsDNA stability, measured as the force of DNAs overstretch transition, increased at low (<10 nM) TMPyP concentrations, then decreased progressively upon increasing TMPyP concentration. The cooperativity of the overstretch transition decreased, due most likely to mechanical roadblocks of ssDNA-bound TMPyP. TMPyP binding increased ssDNAs contour length. The addition of NaCl at high (1 M) concentration competed with many of the nanomechanical changes evoked by TMPyP. Because the largest amplitude of the nanomechanical changes are induced by TMPyP in the pharmacologically relevant nanomolar concentration range, this porphyrin derivative may be used to tune DNAs structure and properties, hence control the myriad of biomolecular processes associated with DNA.

biophysics↗

Temperature-dependent nanomechanics and topography of bacteriophage T7

Viruses are nanoscale infectious agents which may be inactivated by heat treatment. Although heat inactivation is thought to be caused by the release of genetic material from the capsid, the thermally-induced structural changes in viruses are little known. Here we measured the heat-induced changes in the properties of T7 bacteriophage particles exposed to two-stage (65 {degrees}C and 80 {degrees}C) thermal effect by using AFM-based nanomechanical and topographical measurements. We found that exposure to 65 {degrees}C caused the release of genomic DNA due to the loss of the capsid tail which leads to a destabilization of the T7 particles. Further heating to 80 {degrees}C surprisingly led to an increase in mechanical stability due to partial denaturation of the capsomeric proteins kept within the global capsid arrangement.

microbiology↗