Imaging of viruses by atomic force microscopy

Kuznetsov, Yurii G.; Malkin, Alexander J.; Lucas, Robert W.; Plomp, Marco; McPherson, Alexander

doi:10.1099/0022-1317-82-9-2025

Cited by 146 publications

(151 citation statements)

References 25 publications

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“…The images were collected in tapping mode 51,52 at frequencies of about 300 kHz. The forces were similar to those employed in earlier studies on viruses 46,53,54 and biological crystals. [55][56][57] There is considerable previous AFM work concerning double-stranded DNA 49,58,59 but, with some exceptions, 58,[60][61][62] rather little on single-stranded RNA.…”

Section: Afm Proceduressupporting

confidence: 61%

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Atomic Force Microscopy Analysis of Icosahedral Virus RNA

Kuznetsov

Daijogo

Zhou

et al. 2005

Journal of Molecular Biology

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Single-stranded genomic RNAs from four icosahedral viruses (poliovirus, turnip yellow mosaic virus (TYMV), brome mosaic virus (BMV), and satellite tobacco mosaic virus (STMV)) along with the RNA from the helical tobacco mosaic virus (TMV) were extracted using phenol/chloroform. The RNAs were imaged using atomic force microscopy (AFM) under dynamic conditions in which the RNA was observed to unfold. RNAs from the four icosahedral viruses initially exhibited highly condensed, uniform spherical shapes with diameters consistent with those expected from the interiors of their respective capsids. Upon incubation at 26 8C, poliovirus RNA gradually transformed into chains of globular domains having the appearance of thick, irregularly segmented fibers. These ultimately unwound further to reveal segmented portions of the fibers connected by single strands of RNA of 0.5-1 nm thickness. Virtually the same transformations were shown by TYMV and BMV RNA, and with heating, the RNA from STMV. Upon cooling, the chains of domains of poliovirus RNA and STMV RNA condensed and re-formed their original spherical shapes. TMV RNAs initially appeared as single-stranded threads of 0.5-1.0 nm diameter but took on the structure of the multidomain chains upon further incubation at room temperature. These ultimately condensed into short, thick chains of larger domains. Our observations suggest that classical extraction of RNA from icosahedral virions produces little effect on overall conformation. As tertiary structure is lost however, it is evident that secondary structural elements are arranged in a sequential, linear fashion along the polynucleotide chain. At least in the case of poliovirus and STMV, the process of tertiary structure re-formation from the linear chain of secondary structural domains proceeds in the absence of protein. RNA base sequence, therefore, may be sufficient to encode the conformation of the encapsidated RNA even in the absence of coat proteins.

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Section: Afm Proceduressupporting

confidence: 61%

“…Their appearances have been reported elsewhere. 23,39,46,47 Poliovirus was prepared from infected HeLa cells grown to a density of 1.2!10 9 cells/l. The final purification was by centrifugation on 15%-30% (w/v) sucrose gradients at 24 8C for 2.5 hours at 28,000 rpm in a Beckman SW28 swinging bucket rotor.…”

Section: Preparation Of Virusesmentioning

confidence: 99%

Atomic Force Microscopy Analysis of Icosahedral Virus RNA

Kuznetsov

Daijogo

Zhou

et al. 2005

Journal of Molecular Biology

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“…The substrates were then either dried and imaged, or washed two to three times with water before analysis. Fixation with glutaraldehyde has been shown in previous studies not, to the resolution of the AFM technique, to perturb the surface structure of viral particles, and poly-l-lysine insures adherence of cells, virus particles, and their components (Kuznetsov et al, 2001(Kuznetsov et al, , 2002(Kuznetsov et al, , 2003(Kuznetsov et al, , 2004(Kuznetsov et al, , 2005a.…”

Section: Methodsmentioning

confidence: 94%

“…AFM analysis was carried out using a Nanoscope III multimode instrument (Veeco Instruments, Santa Barbara, CA). Imaging procedures were fundamentally the same as described for previous investigations of viruses (Kuznetsov et al, 2001(Kuznetsov et al, , 2002(Kuznetsov et al, , 2003; and RNA (Kuznetsov et al, 2005b;Kuznetsov and McPherson, 2006). For scanning in air, silicon tips were employed.…”

Section: Methodsmentioning

confidence: 99%

An atomic force microscopy investigation of cyanophage structure

Kuznetsov

Chang

Credaroli

et al. 2012

Micron

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“…3) Antibodies to these highly Neuro invasive viruses, Human Herpes Virus 1 & 2, Cytomegalovirus HH5 are found to be the most prevalent and weak. [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23] Cytomegalovirus-infected endothelial cells recruit neutrophils by the secretion of C-X-C chemokines and transmit virus by direct neutrophil-endothelial cell contact during neutrophil transendothelial migration. 32 Therefore, the neuro viral pathology of the heart disrupts the performance of the myocardium and lead to the hypertension and other electrophysiological dysfunctions.…”

Section: Axonal Lesionmentioning

confidence: 99%