Evidence map›Paper›PMID 25320291›Full record

ArticleJournal of virology2015

Nonstructural protein 5A (NS5A) and human replication protein A increase the processivity of hepatitis C virus NS5B polymerase activity in vitro.

Nagraj Mani, Alexander Yuzhakov, Olga Yuzhakov, Joyce T Coll, Jim Black, Kumkum Saxena, John R Fulghum, Judith A Lippke, B Govinda Rao, Rene Rijnbrand and 1 more

Open access · bronzeAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Journal of virology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
0.6field-weighted citation impact, top 32% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

4 citing papers in PubMed, 10 citations in OpenAlex.

  1. Viral Hepatitis - The Road Traveled and the Journey Remaining.Hepatic medicine : evidence and research · 2022
    Review
  2. Review
  3. Review
  4. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

11 authors at 1 institution in 1 country.

Nagraj ManiInfectious Diseases Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA nagraj_mani@yahoo.com.
Alexander YuzhakovInfectious Diseases Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
Olga YuzhakovInfectious Diseases Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
Joyce T CollProtein Sciences Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
Jim BlackProtein Sciences Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
Kumkum SaxenaProtein Sciences Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
John R FulghumProtein Sciences Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
Judith A LippkeProtein Sciences Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
B Govinda RaoComputational Sciences Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
Rene RijnbrandInfectious Diseases Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
Ann D KwongInfectious Diseases Department, Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, USA.
Vertex Pharmaceuticals (United States) · US

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

unlabelledThe precise role(s) and topological organization of different factors in the hepatitis C virus (HCV) RNA replication complex are not well understood. In order to elucidate the role of viral and host proteins in HCV replication, we have developed a novel in vitro replication system that utilizes a rolling-circle RNA template. Under close-to-physiological salt conditions, HCV NS5BΔ21, an RNA-dependent RNA polymerase, has poor affinity for the RNA template. Human replication protein A (RPA) and HCV NS5A recruit NS5BΔ21 to the template. Subsequently, NS3 is recruited to the replication complex by NS5BΔ21, resulting in RNA synthesis stimulation by helicase. Both RPA and NS5A(S25-C447), but not NS5A(S25-K215), enabled the NS5BΔ21-NS3 helicase complex to be stably associated with the template and synthesize RNA product in a highly processive manner in vitro. This new in vitro HCV replication system is a useful tool that may facilitate the study of other replication factors and aid in the discovery of novel inhibitors of HCV replication. IMPORTANCE: The molecular mechanism of hepatitis C virus (HCV) replication is not fully understood, but viral and host proteins collaborate in this process. Using a rolling-circle RNA template, we have reconstituted an in vitro HCV replication system that allows us to interrogate the role of viral and host proteins in HCV replication and delineate the molecular interactions. We showed that HCV NS5A(S25-C447) and cellular replication protein A (RPA) functionally cooperate as a processivity factor to stimulate HCV replication by HCV NS5BΔ21 polymerase and NS3 helicase. This system paves the way to test other proteins and may be used as an assay for discovery of HCV inhibitors.

Indexed as

Host-Pathogen InteractionsVirus ReplicationHepacivirusHumansMutant ProteinsProtein BindingReplication Protein ARNA-Dependent RNA PolymeraseRNA, ViralSequence DeletionViral Nonstructural ProteinsMutant ProteinsNS-5 protein, hepatitis C virusReplication Protein ARNA-Dependent RNA PolymeraseRNA, ViralRPA1 protein, humanViral Nonstructural Proteins

Identifiers

PMID25320291
PMCPMC4301100
OpenAlexW2005406600

What OpenQuestion holds

Textmetadata
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.