Abstract

BackgroundDrosophila Prominin-like is a homolog of mammalian CD133, which is recognized as a biomarker for stem cells. The interacting proteins of CD133 and their biological functions remain elusive.ResultsIn this study, we using yeast two-hybrid assays, GST pull-down assay and co-immunoprecipitation (Co-IP) methods found that Drosophila Prominin-like interacts with ND20, a subunit of mitochondrial respiratory complex I. Bioinformatics analysis suggests that Prominin-like is a six-transmembrane glycoprotein which localizes on cellular membranes. Immunostaining and mitochondrial fractionation indicate that Drosophila Prominin-like could localize in the mitochondria. The knockdown of prominin-like in S2 cells resulted in transient mitochondrial dysfunctions as evidenced by reduced ATP production, elevated ROS generation and an accompanied reduction in mitochondrial proteins. Mitochondrial dysfunctions were detected in aged prominin-like mutant flies.ConclusionOur data indicates that Prominin-like acts to maintain mitochondrial function through its interaction with ND20 which, itself, is active in the mitochondrial electron transport chain. Our study provides insights into a novel molecular mechanism of Drosophila prominin-like and suggests a similar function of CD133 in mammals.

Highlights

  • Drosophila Prominin-like is a homolog of mammalian CD133, which is recognized as a biomarker for stem cells

  • Structural characteristics and the cellular localization of Prominin‐like To evaluate the conservative properties of Promininlike across species, we aligned the amino acid sequences of CD133 from Homo sapiens, Mus musculus, Xenopus tropicalis, Danio rerio with that of Prominin-like from Drosophila melanogaster, using open database from

  • Prominin‐like directly interacts with mitochondrial NADH dehydrogenase 20 kDa subunit (ND20) protein To identify interaction proteins of Prominin-like, we focused on several putative genes including HRB27C, TSG101 (CG9712), and ND20 (CG9172), which were reported in the public data base

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Summary

Introduction

Drosophila Prominin-like is a homolog of mammalian CD133, which is recognized as a biomarker for stem cells. CD133 (AC133/prominin-1) was first identified in 1997 in human hematopoietic progenitor cells and in murine epithelia [1, 2]. It has been widely recognized as a cell-surface marker for stem cells (CSCs) in a variety of tissues and in various cancers including breast carcinomas [3], colon cancer [4], pancreatic cancer [5], hepatocellular carcinoma [6] and neural tumors [7]. Prominin-like has been shown to be localized to the apical protrusions of wing imaginal disc cells, but with unknown function [12].

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