Human cytomegalovirus proteins PP65 and IEP72 are targeted to distinct compartments in nuclei and nuclear matrices of infected human embryo fibroblasts.
The cellular distribution of the human cytomegalovirus (HCMV)-specific UL83 phosphoprotein (pp65) and UL123 immediate-early protein (IEp72) in lytically infected human embryo fibroblasts was studied by means of indirect immunofluorescence and confocal microscopy. Both proteins were found to have a nuclear localization, but they were concentrated in different compartments within the nuclei. The pp65 was located predominantly in the nucleoli; this was already evident with the parental viral protein, which was targeted to the above nuclear compartment very soon after infection. The nucleolar localization of pp65 was also observed at later stages of the HCMV infectious cycle. After chromatin extraction (in the so-called in situ nuclear matrices), a significant portion of the pp65 remained associated with nucleoli within the first hour after infection, then gradually redistributed in a perinucleolar area, as well as throughout the nucleus, with a granular pattern. A quite different distribution was observed for IEp72 at very early stages after infection of human embryo fibroblasts with HCMV; indeed, this viral protein was found in bright foci, clearly observable in both non-extracted nuclei and in nuclear matrices. At later stages of infection, IEp72 became almost homogeneously distributed within the whole nucleus, while the foci increased in size and were more evenly spread; in several infected cells some of them lay within nucleoli. This peculiar nuclear distribution of IEp72 was preserved in nuclear matrices as well. The entire set of data is discussed in terms of the necessity of integration for HCMV-specific products into the pre-existing nuclear architecture, with the possibility of subsequent adaptation of nuclear compartments to fit the needs of the HCMV replicative cycle.
Arcangeletti
M C
MC
Microbiology Section, Department of Pathology and Laboratory Medicine, University of Parma, Parma, Italy. mariacristina.arcangeletti@unipr.it
De Conto
F
F
Ferraglia
F
F
Pinardi
F
F
Gatti
R
R
Orlandini
G
G
Calderaro
A
A
Motta
F
F
Medici
M C
MC
Martinelli
M
M
Valcavi
P
P
Razin
S V
SV
Chezzi
C
C
Dettori
G
G
D002458:Cell Fractionation; D002466:Cell Nucleolus / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism* / Q000648:ultrastructure / Q000821:virology*; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000502:physiology; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism* / Q000648:ultrastructure / Q000821:virology*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D017874:Immediate-Early Proteins / Q000378:metabolism* / Q000648:ultrastructure; D008168:Lung / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D018613:Microscopy, Confocal; D015530:Nuclear Matrix / Q000378:metabolism* / Q000648:ultrastructure / Q000821:virology; D010750:Phosphoproteins / Q000378:metabolism* / Q000648:ultrastructure; D013347:Subcellular Fractions; D014763:Viral Matrix Proteins / Q000378:metabolism* / Q000648:ultrastructure; D014764:Viral Proteins / Q000378:metabolism* / Q000648:ultrastructure; D014779:Virus Replication
C088645:IE1 protein, cytomegalovirus; D017874:Immediate-Early Proteins; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C065418:cytomegalovirus matrix protein 65kDa