Inorganic polyphosphate interacts with nucleolar and glycosomal proteins in trypanosomatids

Molecular Microbiology
Raquel S NegreirosRoberto Docampo

Abstract

Inorganic polyphosphate (polyP) is a polymer of three to hundreds of phosphate units bound by high-energy phosphoanhydride bonds and present from bacteria to humans. Most polyP in trypanosomatids is concentrated in acidocalcisomes, acidic calcium stores that possess a number of pumps, exchangers, and channels, and are important for their survival. In this work, using polyP as bait we identified > 25 putative protein targets in cell lysates of both Trypanosoma cruzi and Trypanosoma brucei. Gene ontology analysis of the binding partners found a significant over-representation of nucleolar and glycosomal proteins. Using the polyphosphate-binding domain (PPBD) of Escherichia coli exopolyphosphatase (PPX), we localized long-chain polyP to the nucleoli and glycosomes of trypanosomes. A competitive assay based on the pre-incubation of PPBD with exogenous polyP and subsequent immunofluorescence assay of procyclic forms (PCF) of T. brucei showed polyP concentration-dependent and chain length-dependent decrease in the fluorescence signal. Subcellular fractionation experiments confirmed the presence of polyP in glycosomes of T. brucei PCF. Targeting of yeast PPX to the glycosomes of PCF resulted in polyP hydrolysis, alteration in their gl...Continue Reading

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Citations

Nov 15, 2018·The Journal of Biological Chemistry·Lihan Xie, Ursula Jakob
May 28, 2019·Life Science Alliance·Laura Lorenzo-OrtsMichael Hothorn
Aug 20, 2019·The New Phytologist·Laura Lorenzo-OrtsMichael Hothorn
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Jan 10, 2021·Current Genetics·Alix Denoncourt, Michael Downey
Jan 14, 2021·Biochemical Society Transactions·Logan P Crowe, Meredith T Morris

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