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Viruses 2014, 6(4), 1612-1636; doi:10.3390/v6041612
Article

Superresolution Imaging of Human Cytomegalovirus vMIA Localization in Sub-Mitochondrial Compartments

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Received: 17 January 2014; in revised form: 16 March 2014 / Accepted: 27 March 2014 / Published: 9 April 2014
(This article belongs to the Special Issue Recent CMV Research)
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Abstract: The human cytomegalovirus (HCMV) viral mitochondria-localized inhibitor of apoptosis (vMIA) protein, traffics to mitochondria-associated membranes (MAM), where the endoplasmic reticulum (ER) contacts the outer mitochondrial membrane (OMM). vMIA association with the MAM has not been visualized by imaging. Here, we have visualized this by using a combination of confocal and superresolution imaging. Deconvolution of confocal microscopy images shows vMIA localizes away from mitochondrial matrix at the Mitochondria-ER interface. By gated stimulated emission depletion (GSTED) imaging, we show that along this interface vMIA is distributed in clusters. Through multicolor, multifocal structured illumination microscopy (MSIM), we find vMIA clusters localize away from MitoTracker Red, indicating its OMM localization. GSTED and MSIM imaging show vMIA exists in clusters of ~100–150 nm, which is consistent with the cluster size determined by Photoactivated Localization Microscopy (PALM). With these diverse superresolution approaches, we have imaged the clustered distribution of vMIA at the OMM adjacent to the ER. Our findings directly compare the relative advantages of each of these superresolution imaging modalities for imaging components of the MAM and sub-mitochondrial compartments. These studies establish the ability of superresolution imaging to provide valuable insight into viral protein location, particularly in the sub-mitochondrial compartments, and into their clustered organization.
Keywords: HCMV vMIA; MAM; mitochondria; OMM; matrix; confocal microscopy; superresolution microscopy; GSTED; MSIM; PALM HCMV vMIA; MAM; mitochondria; OMM; matrix; confocal microscopy; superresolution microscopy; GSTED; MSIM; PALM
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Bhuvanendran, S.; Salka, K.; Rainey, K.; Sreetama, S.C.; Williams, E.; Leeker, M.; Prasad, V.; Boyd, J.; Patterson, G.H.; Jaiswal, J.K.; Colberg-Poley, A.M. Superresolution Imaging of Human Cytomegalovirus vMIA Localization in Sub-Mitochondrial Compartments. Viruses 2014, 6, 1612-1636.

AMA Style

Bhuvanendran S, Salka K, Rainey K, Sreetama SC, Williams E, Leeker M, Prasad V, Boyd J, Patterson GH, Jaiswal JK, Colberg-Poley AM. Superresolution Imaging of Human Cytomegalovirus vMIA Localization in Sub-Mitochondrial Compartments. Viruses. 2014; 6(4):1612-1636.

Chicago/Turabian Style

Bhuvanendran, Shivaprasad; Salka, Kyle; Rainey, Kristin; Sreetama, Sen C.; Williams, Elizabeth; Leeker, Margretha; Prasad, Vidhya; Boyd, Jonathan; Patterson, George H.; Jaiswal, Jyoti K.; Colberg-Poley, Anamaris M. 2014. "Superresolution Imaging of Human Cytomegalovirus vMIA Localization in Sub-Mitochondrial Compartments." Viruses 6, no. 4: 1612-1636.


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