Evidence map›Paper›PMID 34625097›Full record

ArticleParasites & vectors2021

Biotinylation of the Neospora caninum parasitophorous vacuole reveals novel dense granule proteins.

Congshan Yang, Chenrong Wang, Jing Liu, Qun Liu

Open access · goldAbstract read
In one paragraph

Article in Parasites & vectors, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
1.4field-weighted citation impact, top 21% 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

5 citing papers in PubMed, 10 citations in OpenAlex.

  1. Article
  2. TaqMan-quantitative PCR assays applied inFrontiers in cellular and infection microbiology · 2024
    Article
  3. Article
  4. DGPD: a knowledge database of dense granule proteins of the Apicomplexa.Database : the journal of biological databases and curation · 2022
    Article
  5. Review
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

4 authors at 2 institutions in 1 country.

Congshan YangNational Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing, China.
Chenrong WangNational Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing, China.
Jing LiuNational Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing, China.
Qun LiuNational Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing, China. qunliu@cau.edu.cn.
China Agricultural University · CNAnhui Agricultural University · CN

Funding

National Key Research and Development Program of China 2017YFD0501200National Natural Science Foundation of China 31772730Natural Science Foundation of Anhui Province 2008085QC136
6 · The paper itself

Abstract

backgroundNeospora caninum is an obligate intracellular parasite that invades host cells and replicates within the parasitophorous vacuole (PV), which resists fusion with host cell lysosomal compartments. To modify the PV, the parasite secretes an array of proteins, including dense granule proteins (GRAs). The vital role of GRAs in the Neospora life cycle cannot be overestimated. Despite this important role, only a subset of these proteins have been identified, and most of their functions have not been elucidated. Our previous study demonstrated that NcGRA17 is specifically targeted to the delimiting membrane of the parasitophorous vacuole membrane (PVM). In this study, we utilize proximity-dependent biotin identification (BioID) to identify novel components of the dense granules.

methodsNcGRA17 was BirA* epitope-tagged in the Nc1 strain utilizing the CRISPR/Cas9 system to create a fusion of NcGRA17 with the biotin ligase BirA*. The biotinylated proteins were affinity-purified for mass spectrometric analysis, and the candidate GRA proteins from BioID data set were identified by gene tagging. To verify the biological role of novel identified GRA proteins, we constructed the NcGRA23 and NcGRA11 (a-e) knockout strains using the CRISPR/Cas9 system and analyzed the phenotypes of these mutants.

resultsUsing NcGRA17-BirA* fusion protein as bait, we have identified some known GRAs and verified localization of 11 novel GRA proteins by gene endogenous tagging or overexpression in the Nc1 strain. We proceeded to functionally characterize NcGRA23 and NcGRA11 (a-e) by gene knockout. The lack of NcGRA23 or NcGRA11 (a-e) did not affect the parasite propagation in vitro and virulence in vivo.

conclusionsIn summary, our findings reveal that BioID is effective in discovering novel constituents of N. caninum dense granules. The exact biological functions of the novel GRA proteins are yet unknown, but this could be explored in future studies.

Indexed as

AnimalsBiotinBiotinylationFemaleGene Knockout TechniquesLife Cycle StagesMiceMice, Inbred BALB CNeosporaProtozoan ProteinsVacuolesVirulenceBiotinProtozoan ProteinsBioIDCRISPR/Cas9Dense granule proteinNeospora caninumParasitophorous vacuole

Identifiers

PMID34625097
PMCPMC8501707
OpenAlexW3204911789

What OpenQuestion holds

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Registered trials

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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.