{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T03:17:08Z","timestamp":1791429428147,"version":"4.3.3"},"reference-count":46,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2002,4,15]],"date-time":"2002-04-15T00:00:00Z","timestamp":1018828800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Virol"],"published-print":{"date-parts":[[2002,4,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>Assembly of hepatitis delta virus (HDV) in infected human hepatocytes involves association of the 1,679- nucleotide single-stranded genomic RNA (\u03b4RNA) with multiple copies of both small and large forms of the delta protein (\u03b4Ag) to form a ribonucleoprotein particle which in turn interacts with envelope proteins of the natural helper virus, hepatitis B virus. Subsequently, for initiation of a new round of replication, the amount of small \u03b4Ag within the assembled HDV particle is both necessary and sufficient. Quantitative assays were used in order to better understand just how much \u03b4Ag is needed. The molar ratio of \u03b4Ag species to genomic \u03b4RNA in assembled HDV particles was approximately 200. Next, this ratio was determined for cells under several different experimental situations in which HDV genome replication was occurring. These included replication in woodchuck liver and also in mouse liver and skeletal muscle, as well as replication in stably and transiently transfected cultured human hepatoblastoma cells. Surprisingly, in almost all these situations the molar ratios were comparable to that observed for HDV particles. This was true for different times after the initiation of replication and was independent of whether or not virus assembly was occurring. Cell fractionation combined with quantitative assays was used to test whether the majority of \u03b4Ag and \u03b4RNA were colocalized during HDV replication in transfected cells. The cytoplasmic fraction contained the majority of \u03b4Ag and genomic \u03b4RNA. Finally, the quality of \u03b4Ag and \u03b4RNA, especially at relatively late times after the initiation of replication, was examined by using reverse transcription-PCR, cloning, and sequencing through the entire \u03b4Ag open reading frame. When virus assembly and spread were not possible, 20% or less of the predicted \u03b4Ag would have been able to support HDV replication. In summary, an examination of the quantity, quality and intracellular distribution of \u03b4Ag and \u03b4RNA in several different experimental systems has provided a better understanding of the parameters associated with the initiation, maintenance, and ultimate decline of HDV genome replication.<\/jats:p>","DOI":"10.1128\/jvi.76.8.3709-3719.2002","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:59:40Z","timestamp":1027763980000},"page":"3709-3719","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":88,"title":["Parameters of Human Hepatitis Delta Virus Genome Replication: the Quantity, Quality, and Intracellular Distribution of Viral Proteins and RNA"],"prefix":"10.1128","volume":"76","author":[{"given":"Severin","family":"Gudima","sequence":"first","affiliation":[{"name":"Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111-2497"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinhong","family":"Chang","sequence":"additional","affiliation":[{"name":"Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111-2497"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gloria","family":"Moraleda","sequence":"additional","affiliation":[{"name":"Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111-2497"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Azvolinsky","sequence":"additional","affiliation":[{"name":"Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111-2497"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Taylor","sequence":"additional","affiliation":[{"name":"Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111-2497"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","unstructured":"Ausubel F. 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