Temporal expression of SV40 genes occurs in two phases in the lytic cycle. During the early phase, prior to viral DNA replication, early SV40 RNA is synthesized which codes for the early viral T-antigen. Subsequently, late RNA, which codes for the viral structural proteins, is produced in large quantities (5 to 20 times as much as early RNA). The enzymic components responsible for the transition from the early phase to the late phase of the cycle are unclear, but were thought to be linked to viral DNA replication. We have shown that the abundant late transcription depends both on the presence of a functional T-antigen and on an additional factor relating to the onset of DNA replication (perhaps "processing" of the template). Furthermore, we have shown that small amounts of late SV40 RNA can be synthesized prior to DNA replication. This indicates that the transition to predominant late RNA synthesis after DNA replication is a quantitative phenomenon. Using the sarkosyl extraction procedure with incorporation of radiolabeled nucleotides into nascent RNA in vitro, we have shown: 1) Virtually all of the RNA synthesized by the supernatant fraction is virus-specific; 2) qualitatively, the RNA synthesized by the sarkosyl pellet fraction is identical to the RNA synthesized by the supernatant; 3) the rate of synthesis of late: early RNA in an extract obtained late in the lytic cycle is approximately 10:1; 4) there is essentially no initiation of transcription of viral sequences in vitro; 5) the method effectively eliminates processing of viral RNA, allowing for an analysis of nascent transcripts. We have used this method to examine infected cells for potentially unstable mRNAs. We have considered the possibility that human tumors may contain human papovavirus sequences which may or may not be etiologically related to the disease state. It has been shown, for example, that BKV and JCV, two human papovaviruses, are frequently found in immunodeficient patients, the latter case, in the brains of patients with PML. In addition, BKV has been isolated from a brain of a patient with the Wiskott-Aldrich syndrome. We have been screening human tumors and continuous tissue culture lines derived from human tumors as well as normal human tissues for the presence of homologous sequences, and we have examined tumor tissues and sera of cancer patient (Text Truncated -Exceeds Capacity)
NIH ExPorter
NIH ExPorter