M. tuberculosis class II apurinic/ apyrimidinic-endonuclease/3'-5' exonuclease (XthA) engages with NAD+-dependent DNA ligase A (LigA) to counter futile cleavage and ligation cycles in base excision repair.

Class-II AP-endonuclease (XthA) and NAD+-dependent DNA ligase (LigA) are involved in initial and terminal stages of bacterial DNA base excision repair (BER), respectively. XthA acts on abasic sites of damaged DNA to create nicks with 3'OH and 5'-deoxyribose phosphate (5'-dRP) moieties. Co-immunoprecipitation using mycobacterial cell-lysate, identified MtbLigA-MtbXthA complex formation. Pull-down experiments using purified wild-type, and domain-deleted MtbLigA mutants show that LigA-XthA interactions are mediated by the BRCT-domain of LigA. Small-Angle-X-ray scattering, 15N/1H-HSQC chemical shift perturbation experiments and mutational analysis identified the BRCT-domain region that interacts with a novel 104DGQPSWSGKP113 motif on XthA for complex-formation. Isothermal-titration calorimetry experiments show that a synthetic peptide with this sequence interacts with MtbLigA and disrupts XthA-LigA interactions. In vitro assays involving DNA substrate and product analogs show that LigA can efficiently reseal 3'OH and 5'dRP DNA termini created by XthA at abasic sites. Assays and SAXS experiments performed in the presence and absence of DNA, show that XthA inhibits LigA by specifically engaging with the latter's BRCT-domain to prevent it from encircling substrate DNA. Overall, the study suggests a coordinating function for XthA whereby it engages initially with LigA to prevent the undesirable consequences of futile cleavage and ligation cycles that might derail bacterial BER.

Influence of Linker Length on Conformational Preferences of Glycosylated Sugar Amino Acid Foldamers.

Glycosylation of foldamers derived from furanoid sugar amino acids with mannose and a propyltriazole linker results in an unprecedented 16/10 mixed-turn structure in the glycopeptides in water, with a preference for the higher-order structure irrespective of the stereochemistry of the starting foldamer. This is in stark contrast to the structures displayed by the same oligomers in water when mannosylated with a two-carbon-shorter methyltriazole linker: 16-membered turn structure in the cis-foldamer and 10-membered in its trans congener. This demonstrates the defining influence of the linker length on the structural preference of these novel glycopeptide mimics.

Normal adrenal gland: in vivo observations, and high-resolution in vitro chemical shift MR imaging-histologic correlation.

RATIONALE AND OBJECTIVES: The purpose of this study was to determine whether adrenal cortical lipid affects signal intensity on magnetic resonance (MR) images and to evaluate contrast between cortex and medulla. MATERIALS AND METHODS: From their clinical database, the authors selected 37 MR imaging studies of patients with adrenal adenomas. Two independent readers compared in-phase and fat-suppressed T1-weighted images, looking for visible lipid-induced signal intensity loss in the adrenal gland. Six adrenal gland specimens obtained after radical nephrectomy were also studied with high-resolution MR imaging, including in-phase, opposed-phase, and fat-suppressed T1-weighted images, and T2-weighted images. Adjacent histologic sections were stained with oil red O for neutral fats and with hematoxylin-eosin, and they were also viewed with polarization light microscopy. The relative amount of lipid was graded as mild, moderate, or intense, and the appearance of the cortex and medulla was compared with that on the MR images. RESULTS: On the 37 clinical MR studies, there was no visible signal intensity loss within the limbs of the ipsilateral adrenal glands. T2-weighted images of the adrenal specimens showed a thin high-intensity band, corresponding to the appearance of medulla on histologic slices. This could not be seen on any of the T1-weighted images. Region-of-interest measurements were nearly identical for in-phase and opposed-phase images. Histologic analysis showed abundant cortical lipid. CONCLUSION: Adrenal corticomedullary contrast can be depicted on high-resolution T2-weighted images but not on any T1-weighted images. There is abundant cortical lipid in adrenal specimens, but comparison of in-phase with opposed-phase MR images does not depict it.