Dataset Information

Plasma membrane proteome of adhesion-competent endometrial epithelial cells and its modulation by Rab11a

ABSTRACT: Human endometrial epithelium (EE) is composed of a multitude of proteins, amongst which those localized on the plasma membrane (plasma membrane proteins-PMPs) are of critical relevance in early stages of implantation. Evidences support the key role of few PMPs in implantation. However, many remain unidentified, as efforts have not been made till date to generate the plasma membrane proteome of human EE cells, using a gel-free approach. This study presents a protein catalogue of the PMP enriched fraction of Ishikawa cell line; often used as an in vitro model for embryo-adhesive EE. Liquid Chromatography with tandem mass spectrometry identified 3598 proteins. Of these, 1963 proteins were annotated for their membrane localization. Of 1963 proteins, 1321 were found to have transmembrane domain and 43 proteins had GPI anchor. Extensive data mining revealed endometrial expression of 943 proteins reported in human and/or rodents. Further, quantitative alterations were observed in the plasma membrane proteome on perturbation of intracellular trafficking. Silencing of Rab11a (known for its role in plasma membrane organization) expression caused alteration in the abundance of 74 proteins. Caveolin-1 and EpCAM levels were reduced whereas Rab4a abundance increased in the PMP extracts of Rab11a-deficient cells, compared to control cells. Briefly, the study reports the identity of several novel plasma membrane-localized proteins. A major spin-off of the study is the identification of novel proteins trafficked by Rab11a to the plasma membrane. Targeted analysis of novel PMPs may reveal their specific roles in endometrial receptivity and implantation.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: Geetanjali Sachdeva

LAB HEAD: Prof. Geetanjali Sachdeva

PROVIDER: PXD015010 | Pride | 2019-11-08

REPOSITORIES: Pride

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Publications

Plasma membrane proteome of adhesion-competent endometrial epithelial cells and its modulation by Rab11a.

Kakar-Bhanot Ruchi R Brahmbhatt Krupanshi K Kumar Vipin V Suryawanshi Amol R AR Srivastava Sanjeeva S Chaudhari Uddhav U Sachdeva Geetanjali G

Molecular reproduction and development 20191118 1

Human endometrial epithelium (EE) is composed of a multitude of proteins, amongst which those localized on the plasma membrane [plasma membrane proteins (PMPs)] are of critical relevance in the early stages of implantation. Evidence supports the key role of few PMPs in implantation. However, many remain unidentified, as efforts have not been made till date to generate the plasma membrane proteome of human EE cells, using a gel-free approach. This study presents a protein catalog of the PMP enric ...[more]

PMID: 31737970

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Project description:Paramyxoviruses are negative sense single-stranded RNA viruses that comprise many important human and animal pathogens, including human parainfluenza viruses. These viruses bud from the plasma membrane of infected cells after the viral ribonucleoprotein complex (vRNP) is transported from the cytoplasm to the cell membrane via Rab11a marked recycling endosomes. The viral proteins that are critical for mediating this important initial step in viral assembly are unknown. Here we use the model paramyxovirus, murine parainfluenza virus 1, or Sendai virus (SeV), to investigate the roles of viral proteins in Rab11a-driven virion assembly. We previously reported that infection with SeV containing high levels of copy-back defective viral genomes (DVGs) generates heterogenous populations of cells, with cells enriched in full-length virus producing viral particles containing standard or defective viral genomes, while cells enriched in DVGs did not, despite high levels of defective viral genome replication. Here we take advantage of this heterogenous cell phenotype to identify proteins that mediate interaction of vRNPs with Rab11a. We examine the role of matrix protein and nucleoprotein and determine that they are not sufficient to drive interaction of vRNPs and recycling endosomes. Then, using a combination of mass spectrometry and comparative protein abundance and localization in DVG- and FL-high cells, we identify viral polymerase complex components L and, specifically, its cofactor C proteins as interactors with Rab11a. We find that accumulation of these proteins within the cell is the defining feature that differentiates cells that proceed to viral egress from cells which remain in replication phases. Paramyxoviruses are a family of viruses that include a number of pathogens with significant burdens on human health. Particularly, human parainfluenza viruses are an important cause of pneumonia and bronchiolitis in children and do not have any vaccines or direct acting antivirals. These cytoplasmic replicating viruses bud from the plasma membrane and coopt cellular endosomal recycling pathways to traffic viral ribonucleoprotein complexes from the cytoplasm to the membrane of infected cells, yet the viral proteins required for viral engagement with the recycling endosome pathway is not known. Here we use the model paramyxovirus Sendai virus, or murine parainfluenza virus 1, to investigate the role of viral proteins in this initial step in viral assembly. We find that viral polymerase components large protein L and accessory C proteins are necessary for engagement with recycling endosomes. These findings are important in identifying viral targets for the development of antivirals.

Dataset Information

Plasma membrane proteome of adhesion-competent endometrial epithelial cells and its modulation by Rab11a

Publications

Plasma membrane proteome of adhesion-competent endometrial epithelial cells and its modulation by Rab11a.

Similar Datasets

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