Expression of the acidic nuclear immediate-early protein (IE1) of human cytomegalovirus in stable cell lines and its preferential association with metaphase chromosomes.

Virology
R L LafeminaG Hayward

Abstract

Stable DNA-transfected Vero cell lines that express the major immediate-early nuclear antigen (IE68) of HCMV-(Towne) have been established. Immunofluorescence staining with monoclonal antibodies revealed that the protein was distributed either in a uniform diffuse nuclear pattern or as punctate nuclear granules in up to 80% of the cells in these cultures. In addition, 1 to 2% of the positive nuclei gave a distinctive staining pattern suggesting an association with the chromosomes of mitotic cells. Colcemid-blocking studies confirmed that most of the IE antigen was localized in the vicinity of condensed chromosomes in all metaphase cells after methanol fixation. In contrast, the SV40 large T-antigen protein was found to be preferentially excluded from metaphase chromosomes in a similar colcemid-treated human cell line. In transient expression assays, 1 to 2% of IE antigen-positive Vero, 293, or Balb/c3T3 cells also displayed a metaphase chromosome association pattern. Mapping studies using deletion and truncation mutants revealed that the monoclonal antibodies recognized epitopes encoded within the small NH2-terminal exons that are common to both the IE1 and IE2 gene products. However, an intact exon-4 (IE1) region, but not the ...Continue Reading

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Citations

Jul 24, 2013·Proceedings of the National Academy of Sciences of the United States of America·Einat ZalckvarMichael Nevels
Oct 15, 1996·Proceedings of the National Academy of Sciences of the United States of America·E S MocarskiR F Greaves
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Mar 10, 2000·Biochemical and Biophysical Research Communications·J M KimS Kim
Dec 2, 2004·Proceedings of the National Academy of Sciences of the United States of America·Michael NevelsThomas Shenk
Feb 25, 2006·Proceedings of the National Academy of Sciences of the United States of America·Christina PaulusMichael Nevels
Nov 6, 2009·Reviews in Medical Virology·Christina PaulusMichael Nevels
Oct 22, 2002·The Journal of General Virology·Richard StantonGavin W G Wilkinson
Jul 11, 2014·Cell Host & Microbe·Margaret Tarrant-ElorzaGregory S Pari
Apr 25, 2000·Virology·N Bastien, A A McBride
Nov 14, 1997·Journal of Structural Biology·V B BystrevskayaA A Kushch
Jun 7, 2014·Frontiers in Microbiology·Ellen Van Damme, Marnix Van Loock

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