Project summary

Post-translational modification of proteins plays an important role in mediating protein interactions or modulating protein function. Protein ADP-ribosylation is post-translational modification in which an ADP-ribose moiety from NAD+ is transferred to a target protein. The covalent attachment of one ADP-ribose is catalysed by mono-ADP-ribosyl transferases (mARTs) and leads to mono-ADP-ribosylation. The subsequent transfer of additional ADP-ribose molecules is catalysed by poly-ADP-ribose polymerases (PARPs) and results in the synthesis of poly-ADP-ribose (PAR) polymers (long negatively charged chains of repeating ADP-ribose units). Poly-ADP-ribose metabolism is thought to be restricted to eukaryotes, but it has been recently suggested to exist in a scattering of bacterial species. Protein ADP-ribosylation is a reversible modification and it can be removed by the hydrolytic action of mono-ADP-ribose protein hydrolases (mARHs) and poly-ADP-ribose glycohydrolase (PARG).
This project comprises the investigation of reversible protein ADP-ribosylation and its impact on bacterial metabolism, using the genetic model of Streptomyces coelicolor A3(2), an industrially important bacterium from the genus Streptomyces, which produces over two-thirds of all clinically useful antibiotics of natural origin. Bacteria, and Streptomyces in particular are known to utilize protein ADP-ribosylation to control a variety of important pathways (such as morphological differentiation and antibiotic production), but the proteins involved in this regulation are mostly unknown (transferases and hydrolases). The goal of this project is the identification and characterization of the enzymes controlling reversible protein ADP-ribosylation, as well as the pathways and protein targets for their regulation in Streptomyces. In addition, we will further analyse the physiological consequences of the inactivation of the corresponding genes. Our studies are expected to provide an important understanding on how Streptomyces and potentially other bacteria control their metabolism and antibiotic production. Based on genomic data and our preliminary biochemical experiments, we uncovered the SCO6450 protein in Streptomyces as a potential protein responsible for reversing the protein mono-ADP-ribosylation. The conservation of this protein from bacteria to human (SCO6450 orthologue in human is called MacroD1) should allow the extension of our conclusions and analyses to the human model. We will employ a combination of biochemistry and cell biology to understand the function of MacroD1 in human cells.

Project leader

Ivan Ahel, PhD

University of Oxford, Medical Sciences Division, Sir William Dunn School of Pathology

Principal investigator/Project co-leader

Andreja Mikoč, PhD

Ruđer Bošković Institute, Department of Molecular Biology, Laboratory for Molecular Genetics

Team members

Ruđer Bošković Institute, Department of Molecular Biology, Laboratory for Molecular Genetics:
Helena Ćetković, PhD
Dragutin Perina, PhD
Bruna Pleše, PhD
Mirna Imešek, BSc
Helena Bilandžija, PhD
Jasna Lalić, PhD (since March 3^rd, 2014)
Melanija Posavec, PhD (since February 2^nd, 2015)

Ruđer Bošković Institute, Division of Physical Chemistry, Laboratory for chemical and biological crystallography:
Marija Luić, PhD
Igor Sabljić, BSc
Zoran Štefanić, PhD
Ivana Leščić Ašler, PhD

Ruđer Bošković Institute, Division for Marine and Environmental Research, Laboratory for Analytical Chemistry and Biogeochemistry of Organic Compounds:
Marijan Ahel, PhD

University of Oxford, Medical Sciences Division, Sir William Dunn School of Pathology:
Roko Žaja, PhD

Collaborators

Dr. Giselda Bucca, University of Surrey, Guildford, UK
Dr. Ivan Matić, Max Planck Institute for Biology of Ageing, Cologne, Germany

Publications

Perina D, Mikoč A, Ahel J, Ćetković H, Žaja R, Ahel I. (2014) Distribution of protein poly(ADP-ribosyl)ation systems across all domains of life. DNA Repair (Amst). 23:4-16.

Perina D, Korolija M, Hadžija MP, Grbeša I, Belužić R, Imešek M, Morrow C, Marjanović MP, Bakran-Petricioli T, Mikoč A, Ćetković H. (2015) Functional and structural characterization of FAU gene/protein from marine sponge Suberites domuncula. Mar Drugs. 13:4179-96.

Conferences

Jasna Lalić, Andreja Mikoč, Dragutin Perina, Igor Sabljić, Bruna Pleše, Mirna Imešek, Helena Ćetković , Marija Luić, Roko Žaja, Ivan Ahel: Macrodomain protein from bacterium Streptomyces coelicolor, Congress of the Croatian Society of Biochemistry and Molecular Biology: The Interplay of Biomolecules. Zadar, Croatia; 24-27 September 2014 (poster presentation)

Dragutin Perina, Andreja Mikoč, Josip Ahel, Helena Ćetković, Roko Žaja, Ivan Ahel: Poly(ADP-ribosyl)ation in the red seaweed Chondrus crispus, Congress of the Croatian Society of Biochemistry and Molecular Biology: The Interplay of Biomolecules, Zadar, Croatia; 24-27 September 2014 (poster presentation)

Dragutin Perina, Andreja Mikoč, Josip Ahel, Helena Ćetković, Roko Žaja, Ivan Ahel: Distribution of protein poly(ADP-ribosyl)ation systems across all domains of life, OMICs in Biomedical Research, MedILS, Split, Croatia; 8-12 June 2015 (poster presentation)

Ivan Ahel: Enzymes and mechanisms involved in the reversal of protein ADP-ribosylation, FASEB Conference: NAD Metabolism and Signaling, Timmendorfer Strand, Germany; 9-14 August, 2015 (invited lecture)

Jasna Lalić, Melanija Posavec Marjanović, Dragutin Perina, Igor Sabljić, Bruna Pleše, Mirna Imešek, Roko Žaja, Helena Ćetković, Marija Luić, Andreja Mikoč, Ivan Ahel: Macrodomain protein from bacterium Streptomyces coelicolor, FASEB Conference: NAD Metabolism and Signaling, Timmendorfer Strand, Germany; 9-14 August, 2015 (poster presentation)

Dragutin Perina, Marina Korolija, Marijana Popović Hadžija, Ivana Grbeša, Robert Belužić, Mirna Imešek, Christine Morrow, Tatjana Bakran-Petricioli, Andreja Mikoč, Helena Ćetković,: Characterization of Fau gene and protein from marine sponge Suberites domuncula, 12^th Croatian Biological Congress, Sveti Martin na Muri, Croatia; 18-23 September, 2015 (poster presentation)

Dragutin Perina, Andreja Mikoč, Josip Ahel, Helena Ćetković, Roko Žaja, Ivan Ahel: Distribution of protein poly(ADP-ribosyl)ation systems across all domains of life, 12^th Croatian Biological Congress, Sveti Martin na Muri, Croatia; 18-23 September, 2015 (poster presentation)

Project activities