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. 1994 Feb;66(2 Pt 1):293–298. doi: 10.1016/s0006-3495(94)80800-8

Structures of large T antigen at the origin of SV40 DNA replication by atomic force microscopy.

I A Mastrangelo 1, M Bezanilla 1, P K Hansma 1, P V Hough 1, H G Hansma 1
PMCID: PMC1275694  PMID: 8161681

Abstract

For inorganic crystals such as calcite (CaCO3), Atomic Force Microscopy (AFM) has provided surface structure at atomic resolution (Ohnesorge and Binnig, 1993). As part of a broad effort to obtain high resolution for an individual protein or protein assembly (Binnig et al., 1986; Rugar and Hansma, 1990; Radmacher et al., 1992), we applied AFM to study the ATP-dependent double hexamer of SV40 large T antigen, which assembles around the viral origin of DNA replication. Multimeric mass has been determined in two-dimensional projected images by Scanning Transmission Electron Microscopy (STEM) (Mastrangelo et al., 1989). By AFM, if the DNA-protein preparation has been stained positively by uranyl acetate, the contour at the junction between hexamers is visible as a cleft, 2-4 nm deep. The cleft, whether determined as a fraction of height by AFM or as a fraction of mass thickness by STEM, is of comparable magnitude. On either side of the cleft, hexamers attain a maximum height of 13-16 nm. Monomers found in the absence of ATP show heights of 5-7 nm. Taken together, the z coordinates provide a surface profile of complete and partial replication assemblies consistent with the spatial distribution of recognition pentanucleotides on the DNA, and they contribute direct geometrical evidence for a ring-like hexamer structure.

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295FIGURE 1
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296FIGURE 2
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297FIGURE 3
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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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