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Comparative analysis of MACROD1, MACROD2 and TARG1 expression, localisation and interactome
R Žaja1, G Aydin2, B E Lippok2
1Institute of Biochemistry and Molecular Biology, RWTH Aachen University, Pauwelsstrasse 30, 52074, Aachen, Germany. rzaja@ukaachen.de.
Scientific Reports
|May 20, 2020
Summary
Researchers developed new antibodies to study MACROD1, MACROD2, and TARG1 enzymes, which reverse mono(ADP-ribosyl)ation (MARylation). Their distinct cellular locations and functions, particularly in mitochondria and nucleoli, were revealed, offering insights into RNA metabolism.
Area of Science:
- Biochemistry and Molecular Biology
- Cellular Biology
- Posttranslational Modifications
Background:
- Mono(ADP-ribosyl)ation (MARylation) is a reversible posttranslational modification crucial for cellular processes.
- MACROD1, MACROD2, and TARG1 are macrodomain-containing enzymes that reverse MARylation in vitro.
- The distinct cellular roles of these hydrolases remain unclear due to limited validated reagents and contradictory data.
Purpose of the Study:
- To develop validated monoclonal antibodies for studying MACROD1, MACROD2, and TARG1.
- To determine the tissue distribution and intracellular localization of these MARylation hydrolases.
- To investigate the cellular functions and interactomes of MACROD1 and TARG1.
Main Methods:
- Development and application of monoclonal antibodies for protein analysis.
- Tissue distribution and immunofluorescence microscopy for cellular localization.
- BioID proximity labeling for interactome mapping.
- Gene Ontology (GO) analysis of identified interactors.
Main Results:
- MACROD1 localizes to mitochondria in skeletal muscle; MACROD2 is found in neuroblastoma cells (nucleo- and cytoplasm); TARG1 is ubiquitously expressed (nucleoplasm, nucleolus, stress granules).
- Loss of MACROD1 or TARG1 disrupts mitochondrial or nucleolar morphology, respectively.
- Interactome analysis supports compartment-specific localization and suggests roles in RNA metabolism for MACROD1 and TARG1.
Conclusions:
- Validated antibodies enabled detailed characterization of MARylation hydrolase expression, localization, and interactions.
- MACROD1 and TARG1 play distinct, organelle-specific roles, particularly in mitochondrial and nucleolar function and RNA metabolism.
- This study provides a foundation for further research into the physiological functions of these key enzymes.
