DNA Ligase C and Prim-PolC participate in base excision repair in mycobacteria

Prokaryotic Ligase D is a conserved DNA repair apparatus processing DNA double-strand breaks in stationary phase. An orthologous Ligase C (LigC) complex also co-exists in many bacterial species but its function is unknown. Here, we show that the LigC complex interacts with core BER enzymes in vivo and demonstrate that together these factors constitute an excision repair apparatus capable of repairing damaged bases and abasic sites. The polymerase component, which contains a conserved C-terminal structural loop, preferentially binds to and fills-in short gapped DNA intermediates with RNA and LigC ligates the resulting nicks to complete repair. Components of the LigC complex, like LigD, are expressed upon entry into stationary phase and cells lacking either of these pathways exhibit increased sensitivity to oxidising genotoxins. Together, these findings establish that the LigC complex is directly involved in an excision repair pathway(s) that repairs DNA damage with ribonucleotides during stationary phase

Molecular basis for DNA repair synthesis on short gaps by mycobacterial Primase-Polymerase C

Cells utilise specialized polymerases from the Primase-Polymerase (Prim-Pol) superfamily to maintain genome stability. Prim-Pol’s function in genome maintenance pathways including replication, repair and damage tolerance. Mycobacteria contain multiple Prim-Pols required for lesion repair, including Prim-PolC that performs short gap repair synthesis during excision repair. To understand the molecular basis of Prim-PolC’s gap recognition and synthesis activities, we elucidated crystal structures of pre- and post-catalytic complexes bound to gapped DNA substrates. These intermediates explain its binding preference for short gaps and reveal a distinctive modus operandi called Synthesis-dependent Template Displacement (STD). This mechanism enables Prim-PolC to couple primer extension with template base dislocation, ensuring that the unpaired templating bases in the gap are ushered into the active site in an ordered manner. Insights provided by these structures establishes the molecular basis of Prim-PolC’s gap recognition and extension activities, while also illuminating the mechanisms of primer extension utilised by closely related Prim-Pols

Synthesis and decomposition mechanisms of ternary Mg2COH5 studied using in situ synchrotron X-ray diffraction

A ternary Mg2COH5 hydride was synthesized using a novel method that relies on a relatively short mechanical milling time (1 h) of a 2:1 MgH2-Co powder mixture followed by sintering at a sufficiently high hydrogen pressure (>85 bar) and heating from RT to 500 degrees C. The ternary hydride forms in less than 2.5 h (including the milling time) with a yield of similar to 90% at similar to 300 degrees C. The mechanisms of formation and decomposition of ternary Mg2COH5 were studied in detail using an in situ synchrotron radiation powder X-ray diffraction (SR-PXD). The obtained experimental results are supported by morphological and microstructural investigations performed using SEM and high-resolution STEM. Additionally, thermal effects occurring during the desorption reaction were studied using DSC. The morphology of as-prepared ternary Mg2COH5 is characterized by the presence of porous particles with various shapes and sizes, which, in fact, are a type of nanocomposite consisting mainly of nanocrystallites with a size of similar to 5 nm. Mg2COH5 decomposes at approximately 300 degrees C to elemental Mg and Co. Additionally, at approximately 400 degrees C, MgCo is formed as precipitates inserted into the Mg Co matrix. During the rehydrogenation of the decomposed residues, prior to the formation of Mg2COH5, MgH2 appears, which confirms its key role in the synthesis of the ternary Mg2COH5. Copyright (C) 2011, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved

Mg2NiH4 synthesis and decomposition reactions

A ternary Mg2NiH4 hydride was synthesized using method that relies on a relatively short mechanical milling time (one hour) of a 2:1 MgH2-Ni powder mixture followed by sintering at a sufficiently high hydrogen pressure (>85 bar) and temperature (>400 degrees C). The ternary hydride forms in less than 2.5 h (including the milling time) with a yield of similar to 90% as a mixture of two polymorphic forms. The mechanisms of formation and decomposition of ternary Mg2NiH4 under different hydrogen pressures were studied in detail using an in situ synchrotron radiation powder X-ray diffraction (SR-PXD) and high pressure DSC. The obtained experimental results are supported by morphological and microstructural investigations performed using SEM and high resolution STEM. Additionally, effects occurring during the desorption reaction were studied using DSC coupled with mass spectrometry. Copyright (C) 2013, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved