Abstract. The PML protein was first identified as part of a fusion product with the retinoic acid receptor (RARa), resulting from the t(15;17) chromosomal translocation associated with acute promyelocytic leukemia (APL). It has been previously demonstrated that PML, which is tightly bound to the nuclear matrix, concentrates in discrete subnuclear compartments that are disorganized in APL cells due to the expression of the PML-RARa hybrid. Here we report that adenovirus infection causes a drastic redistribution of PML from spherical nuclear bodies into fibrous structures. The product encoded by adenovirus E4-ORF3 is shown to be responsible for this reorganization and to colocalize with PML into these fibers. In addition, we demonstrate that E1A oncoproteins concentrate in the PML domains, both in infected and transiently transfected cells, and that this association requires the conserved amino acid motif (D)LXCXE, common to all viral oncoproteins that bind pRB or the related p107 and p130 proteins. The SV-40 large T antigen, another member of this oncoprotein family is also found in close association with the PML nuclear bodies. Taken together, the present data indicate that the subnuclear domains containing PML represent a preferential target for DNA tumor viruses, and therefore suggest a more general involvement of the PML nuclear bodies in oncogenic processes.T HE eukaryotic nucleus is highly organized into discrete domains which spatially separate different biochemical processes. A variety of metabolic activities such as DNA replication, ribosome assembly, transcription, and pre-mRNA processing localize to distinct subnuclear compartments. The most conspicuous example is the nucleolus in which rRNAs are assembled into ribosomal subunits (reviewed by Scheer and Weisenberger, 1994). DNA replication sites were also shown to concentrate within discrete regions containing the proliferating cell nuclear antigen (PCNA; 1 Bravo and MacdonaldBravo, 1987) as well as DNA methyltransferase (Leonhardt et al., 1992). In addition, the small nuclear ribonucleoprotein particles (snRNPs), which are the major subunits of spliceosomes, were similarly found to localize to T. Carvalho and J.-S. Seeler should be considered as first authors.

All mRNAs expressed from the adenovirus major late transcription unit have a common, 201-nucleotide-long 5' leader sequence, which consists of three short exons (the tripartite leader). This leader has two

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Here we show that the adenovirus early region 4 (E4) open reading frame 4 (ORF4) protein autoregulates its own transcription by inhibiting adenovirus E1A-induced activation of E4 transcription both in transient transfection experiments and during lytic virus growth. The inhibitory activity of E4-ORF4 was selective for E1A-CR3-dependent transactivation and had no effect on CR1 transactivation. The inhibitory activity of E4-ORF4 was relieved by okadaic acid treatment, which inhibits the cellular protein phosphatase 2A (PP2A), suggesting that E4-ORF4 controls the phosphorylated status of transcription factors important for E4 promoter activity. This conclusion agrees with previous demonstrations that E4-ORF4 associates with PP2A and causes a partial dephosphorylation of certain transcription factors, including E1A (U. Müller, T. Kleinberger, and T.

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