PMID- 2477948
OWN - NLM
STAT- MEDLINE
DCOM- 19891107
LR  - 20190714
IS  - 0042-6822 (Print)
IS  - 0042-6822 (Linking)
VI  - 172
IP  - 2
DP  - 1989 Oct
TI  - Expression of the acidic nuclear immediate-early protein (IE1) of human 
      cytomegalovirus in stable cell lines and its preferential association with 
      metaphase chromosomes.
PG  - 584-600
AB  - 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 exon-5 (IE2) region 
      of the HCMV IE gene complex, was required for conferring both the normal diffuse 
      nuclear localization pattern and the chromosome-association properties. 
      Furthermore, removal of the glutamic acid-rich COOH-terminal coding portions of 
      exon-4 resulted in aberrant staining patterns with production of large, 
      phase-dense nuclear globules in all positive cells. An association between the 
      IE68 IE1 protein and metaphase chromosomes was also detected after HCMV-(Towne) 
      infection in a small proportion of both nonpermissive Balb/c3T3 cells and 
      permissive HF cells. We conclude that the IE1 acidic nuclear phosphoprotein 
      displays some properties similar to those of the EBNA-1 protein of Epstein-Barr 
      virus and suggest that it may potentially play a role in maintenance of the 
      latent state of HCMV DNA.
FAU - Lafemina, R L
AU  - Lafemina RL
AD  - Department of Pharmacology and Molecular Sciences, Johns Hopkins University 
      School of Medicine, Baltimore, Maryland 21205.
FAU - Pizzorno, M C
AU  - Pizzorno MC
FAU - Mosca, J D
AU  - Mosca JD
FAU - Hayward, G S
AU  - Hayward GS
LA  - eng
GR  - R01 AI24576/AI/NIAID NIH HHS/United States
GR  - R01 CA28473/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Virology
JT  - Virology
JID - 0110674
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antigens, Viral)
RN  - 0 (Epitopes)
RN  - 0 (Immediate-Early Proteins)
RN  - 0 (immediate-early proteins, cytomegalovirus)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal/immunology
MH  - Antigens, Viral/biosynthesis/*genetics
MH  - Cell Line
MH  - Chromosomes/*analysis
MH  - Cytomegalovirus/*genetics/immunology
MH  - Epitopes/analysis
MH  - Fluorescent Antibody Technique
MH  - Humans
MH  - *Immediate-Early Proteins
MH  - *Metaphase
MH  - Plasmids
MH  - Transfection
MH  - Vero Cells
EDAT- 1989/10/01 00:00
MHDA- 1989/10/01 00:01
CRDT- 1989/10/01 00:00
PHST- 1989/10/01 00:00 [pubmed]
PHST- 1989/10/01 00:01 [medline]
PHST- 1989/10/01 00:00 [entrez]
AID - 0042-6822(89)90201-8 [pii]
AID - 10.1016/0042-6822(89)90201-8 [doi]
PST - ppublish
SO  - Virology. 1989 Oct;172(2):584-600. doi: 10.1016/0042-6822(89)90201-8.