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

Viruses

Shivaprasad Bhuvanendran³  Kyle Salka³  Kristin Rainey¹  Sen Chandra Sreetama³  Elizabeth Williams³  Margretha Leeker³  Vidhya Prasad³  Jonathan Boyd²  George H. Patterson¹  Jyoti K. Jaiswal³ 

[1] Section on Biophotonics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20892, USA; E-Mail:;Life Science Division, Leica Microsystems, Inc., 1700 Leider Lane, Buffalo Grove, IL 60089, USA; E-Mail:;Research Center for Genetic Medicine, Children’s Research Institute, Children’s National Health System, 111 Michigan Avenue, NW, Washington, DC 20010, USA; E-Mails:

关键词: HCMV vMIA;    MAM;    mitochondria;    OMM;    matrix;    confocal microscopy;    superresolution microscopy;    GSTED;    MSIM;    PALM;   

DOI  :  10.3390/v6041612

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.