{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T02:14:14Z","timestamp":1789352054548,"version":"build-2803163510"},"reference-count":48,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2016,4,15]],"date-time":"2016-04-15T00:00:00Z","timestamp":1460678400000},"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":[[2016,4,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Adeno-associated virus (AAV) has long been known to inhibit helper adenovirus (Ad) replication independently of AAV Rep protein expression. More recently, replication of Ad serotype 5 (Ad5)\/AAV serotype 2 (AAV-2) hybrid vectors was shown to be inhibited in\n                    <jats:italic>cis<\/jats:italic>\n                    by a sequence near the 3\u2032 end of AAV\n                    <jats:italic>rep<\/jats:italic>\n                    , termed the Rep inhibition sequence for adenoviral replication (RIS-Ad). RIS-Ad functions independently of Rep protein expression. Here we demonstrate that inhibition of adenoviral replication by RIS-Ad requires an active AAV p40 promoter and the 5\u2032 half of the intron. In addition, Ad inhibition is critically dependent on the integrity of the p40 transcription start site (TSS) leading to short p40-associated transcripts. These do not give rise to effector molecules capable of inhibiting adenoviral replication in\n                    <jats:italic>trans<\/jats:italic>\n                    , like small polypeptides or microRNAs. Our data point to an inhibitory mechanism in which RNA polymerase II (Pol II) pauses directly downstream of the p40 promoter, leading to interference of the stalled Pol II transcription complex with the adenoviral replication machinery. Whereas inhibition by RIS-Ad is mediated exclusively in\n                    <jats:italic>cis<\/jats:italic>\n                    , it can be overcome by providing a replication-competent adenoviral genome in\n                    <jats:italic>trans<\/jats:italic>\n                    . Moreover, the inhibitory effect of RIS-Ad is not limited to AAV-2 but could also be shown for the corresponding regions of other AAV serotypes, including AAV-5. These findings have important implications for the future generation of Ad5\/AAV hybrid vectors.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>IMPORTANCE<\/jats:bold>\n                    Insertion of sequences from the 3\u2032 part of the\n                    <jats:italic>rep<\/jats:italic>\n                    gene of adeno-associated virus (AAV) into the genome of its helper adenovirus strongly reduces adenoviral genome replication. We could show that this inhibition is mediated exclusively in\n                    <jats:italic>cis<\/jats:italic>\n                    without the involvement of\n                    <jats:italic>trans<\/jats:italic>\n                    -acting regulatory RNAs or polypeptides but nevertheless requires an active AAV-2 p40 promoter and p40-associated short transcripts. Our results suggest a novel inhibitory mechanism that has so far not been described for AAV and that involves stalled RNA polymerase II complexes and their interference with adenoviral DNA replication. Such a mechanism would have important implications both for the generation of adenoviral vectors expressing the AAV\n                    <jats:italic>rep<\/jats:italic>\n                    and\n                    <jats:italic>cap<\/jats:italic>\n                    genes and for the regulation of AAV gene expression in the absence and presence of helper virus.\n                  <\/jats:p>","DOI":"10.1128\/jvi.03120-15","type":"journal-article","created":{"date-parts":[[2016,2,3]],"date-time":"2016-02-03T22:38:55Z","timestamp":1454539135000},"page":"3981-3993","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":7,"title":["A Regulatory Element Near the 3\u2032 End of the Adeno-Associated Virus\n                    <i>rep<\/i>\n                    Gene Inhibits Adenovirus Replication in\n                    <i>cis<\/i>\n                    by Means of p40 Promoter-Associated Short Transcripts"],"prefix":"10.1128","volume":"90","author":[{"given":"Stefan","family":"Weger","sequence":"first","affiliation":[{"name":"Institute of Virology, Charit\u00e9-University Medicine Berlin, Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Hammer","sequence":"additional","affiliation":[{"name":"Institute of Virology, Charit\u00e9-University Medicine Berlin, Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melanie","family":"Gonsior","sequence":"additional","affiliation":[{"name":"Institute of Virology, Charit\u00e9-University Medicine Berlin, Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catrin","family":"Stutika","sequence":"additional","affiliation":[{"name":"Institute of Virology, Charit\u00e9-University Medicine Berlin, Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Regine","family":"Heilbronn","sequence":"additional","affiliation":[{"name":"Institute of Virology, Charit\u00e9-University Medicine Berlin, Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.149.3685.754"},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.40.1.241-247.1981"},{"key":"e_1_3_1_4_2","first-page":"1","article-title":"Biology of adeno-associated virus","volume":"218","author":"Berns KI","year":"1996","unstructured":"Berns KI, Giraud C. 1996. 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