Ability of minus strands and modified plus strands to act as templates in Semliki Forest virus RNA replication.

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Sammanfattning

During virus multiplication, the viral genome is recognized and recruited for replication based on specific cis-acting elements. Here we have dissected the important cis-acting sequence elements in Semliki Forest virus RNA, by using a trans-replication system. Since the viral replicase is expressed from a separate plasmid, the template RNA can be freely modified in this system. We show that the cis-acting element at the beginning of the non-structural protein 1 (nsP1) coding region together with the end of the 3′ untranslated region are the minimal requirements for minus strand synthesis. To achieve a high level of replication, the native 5′ untranslated region was also needed. The virus-induced membranous replication compartments or spherules were only detected when a replication-competent template was present with an active replicase and minus strands were produced. No translation could be detected from the minus strands, suggesting that they are segregated from the cytoplasm. Minus strands could not directly be recruited to initiate the replication process. Thus, there is only one defined pathway for replication, starting with plus strand recognition followed by concomitant spherule formation and minus strand synthesis.
Originalspråkengelska
TidskriftJournal of General Virology
Volym87
Utgåva6
Sidor (från-till)1395-1407
Antal sidor13
ISSN0022-1317
DOI
StatusPublicerad - 2016
MoE-publikationstypA1 Tidskriftsartikel-refererad

Vetenskapsgrenar

  • 1183 Växtbiologi, mikrobiologi, virologi

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title = "Ability of minus strands and modified plus strands to act as templates in Semliki Forest virus RNA replication.",
abstract = "During virus multiplication, the viral genome is recognized and recruited for replication based on specific cis-acting elements. Here we have dissected the important cis-acting sequence elements in Semliki Forest virus RNA, by using a trans-replication system. Since the viral replicase is expressed from a separate plasmid, the template RNA can be freely modified in this system. We show that the cis-acting element at the beginning of the non-structural protein 1 (nsP1) coding region together with the end of the 3′ untranslated region are the minimal requirements for minus strand synthesis. To achieve a high level of replication, the native 5′ untranslated region was also needed. The virus-induced membranous replication compartments or spherules were only detected when a replication-competent template was present with an active replicase and minus strands were produced. No translation could be detected from the minus strands, suggesting that they are segregated from the cytoplasm. Minus strands could not directly be recruited to initiate the replication process. Thus, there is only one defined pathway for replication, starting with plus strand recognition followed by concomitant spherule formation and minus strand synthesis.",
keywords = "1183 Plant biology, microbiology, virology",
author = "Hellstr{\"o}m, {Kirsi Johanna} and Kallio, {Katri Anneli} and Hanna-Mari Meril{\"a}inen and Jokitalo, {Eija Sofia} and Ahola, {Tero Tapani}",
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T1 - Ability of minus strands and modified plus strands to act as templates in Semliki Forest virus RNA replication.

AU - Hellström, Kirsi Johanna

AU - Kallio, Katri Anneli

AU - Meriläinen, Hanna-Mari

AU - Jokitalo, Eija Sofia

AU - Ahola, Tero Tapani

PY - 2016

Y1 - 2016

N2 - During virus multiplication, the viral genome is recognized and recruited for replication based on specific cis-acting elements. Here we have dissected the important cis-acting sequence elements in Semliki Forest virus RNA, by using a trans-replication system. Since the viral replicase is expressed from a separate plasmid, the template RNA can be freely modified in this system. We show that the cis-acting element at the beginning of the non-structural protein 1 (nsP1) coding region together with the end of the 3′ untranslated region are the minimal requirements for minus strand synthesis. To achieve a high level of replication, the native 5′ untranslated region was also needed. The virus-induced membranous replication compartments or spherules were only detected when a replication-competent template was present with an active replicase and minus strands were produced. No translation could be detected from the minus strands, suggesting that they are segregated from the cytoplasm. Minus strands could not directly be recruited to initiate the replication process. Thus, there is only one defined pathway for replication, starting with plus strand recognition followed by concomitant spherule formation and minus strand synthesis.

AB - During virus multiplication, the viral genome is recognized and recruited for replication based on specific cis-acting elements. Here we have dissected the important cis-acting sequence elements in Semliki Forest virus RNA, by using a trans-replication system. Since the viral replicase is expressed from a separate plasmid, the template RNA can be freely modified in this system. We show that the cis-acting element at the beginning of the non-structural protein 1 (nsP1) coding region together with the end of the 3′ untranslated region are the minimal requirements for minus strand synthesis. To achieve a high level of replication, the native 5′ untranslated region was also needed. The virus-induced membranous replication compartments or spherules were only detected when a replication-competent template was present with an active replicase and minus strands were produced. No translation could be detected from the minus strands, suggesting that they are segregated from the cytoplasm. Minus strands could not directly be recruited to initiate the replication process. Thus, there is only one defined pathway for replication, starting with plus strand recognition followed by concomitant spherule formation and minus strand synthesis.

KW - 1183 Plant biology, microbiology, virology

U2 - 10.1099/jgv.0.000448

DO - 10.1099/jgv.0.000448

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JO - Journal of General Virology

JF - Journal of General Virology

SN - 0022-1317

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