HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 286/6/C1324    most recent 00404.2003v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (3) Citing Articles via Google Scholar Google Scholar Articles by Maglova, L. M. Articles by Russell, J. M. Search for Related Content PubMed PubMed Citation Articles by Maglova, L. M. Articles by Russell, J. M.

MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

Perinuclear localization of Na-K-Cl-cotransporter protein after human cytomegalovirus infection

Biological Research Laboratories, Syracuse University, Syracuse, New York 13244

Submitted 22 September 2003 ; accepted in final form 16 January 2004

We (41) previously reported that Na-K-Cl-cotransporter (NKCC) function and microsomal protein expression are both dramatically reduced late in human cytomegalovirus (HCMV) infection of a human fibroblast cell line (MRC-5). We now report DNA microarray data showing that no significant HCMV-dependent NKCC gene repression can be detected 30 h postexposure (PE) to the virus. Consequently, we used plasma membrane biotinylation and subsequent subcellular fractionation in combination with semiquantitative immunoblotting and confocal microscopy to investigate the possibility that intracellular redistribution of the NKCC protein after HCMV infection could be a cause of the HCMV-induced loss of NKCC ion transport function. Our results show that the lifetime of plasmalemmal NKCC protein in quiescent, uninfected MRC-5 cells is 48 h, and <20% of the total expressed NKCC protein are in the plasma membrane. The remainder (80%) was detected as diffusely distributed, small punctate structures in the cytoplasm. Following HCMV infection: 1) NKCC protein expression in the plasmalemma was sharply reduced (75%) within 24 h PE and thereafter continued to slowly decrease; 2) total cellular NKCC protein content remained unchanged or slightly increased during the course of the viral infection; and 3) HCMV infection caused NKCC protein to accumulate in the perinuclear region late in the HCMV infection (72 h PE). Thus our results imply that, in the process of productive HCMV infection, NKCC protein continues to be synthesized, but, instead of being delivered to the plasma membrane, it is clustered in a large, detergent-soluble perinuclear structure.

sodium-potassium-chloride-cotransporter; human fibroblast cell line; perinuclear accumulation

This article has been cited by other articles:

				W. E. Crowe, L. M. Maglova, P. Ponka, and J. M. Russell Human cytomegalovirus-induced host cell enlargement is iron dependent Am J Physiol Cell Physiol, October 1, 2004; 287(4): C1023 - C1030. [Abstract] [Full Text] [PDF]