DNA-dependent SUMO modification of PARP-1.

Poly(ADP-ribose) polymerase 1 (PARP-1) plays an important role in DNA repair, but also contributes to other aspects of nucleic acid metabolism, such as transcriptional regulation. Modification of PARP-1 with the small ubiquitin-related modifier (SUMO) affects its function as a transcriptional co-activator of hypoxia-responsive genes and promotes induction of the heat shock-induced HSP70.1 promoter. We now report that PARP-1 sumoylation is strongly influenced by DNA. Consistent with a function in transcription, we show that sumoylation in vitro is enhanced by binding to intact, but not to damaged DNA, in a manner clearly distinct from the mechanism by which DNA damage stimulates PARP-1's catalytic activity. An enhanced affinity of PARP-1 for the SUMO-conjugating enzyme Ubc9 upon binding to DNA is likely responsible for this effect. Sumoylation does not interfere with the catalytic or DNA-binding properties of PARP-1, and structural analysis reveals no significant impact of SUMO on the conformation of PARP-1's DNA-binding domain. In vivo, sumoylated PARP-1 is associated with chromatin, but the modification is not responsive to DNA damage and is not affected by PARP-1 catalytic activity. Our results suggest that PARP-1's alternative modes of DNA recognition serve as a means to differentiate between distinct aspects of the enzyme's function.

Posterior periosteal disruption in Salter-Harris Type II fractures of the distal femur: evidence for a hyperextension mechanism.

OBJECTIVE: Patterns of periosteal disruption are important factors in assessing the mechanism of injury of radiologically evident Salter-Harris (SH) fractures. The purpose of this study is to assess the frequency of posterior periosteal disruption on MRI in radiographically occult or subtle SH type II fractures of the distal femur and to evaluate associated soft-tissue findings that support a hyperextension mechanism of injury. CONCLUSION: We found that all children in our experience with occult or subtle SH type II fractures of the distal femur have posterior periosteal disruption and other MRI findings to indicate a hyperextension mechanism of injury. Direct indicators of fracture may be inconspicuous, and the presence of posterior periosteal disruption is a clue that should prompt a search for other features of this serious pediatric injury, which may be followed by limb shortening or angular deformity.

RAD51C is required for Holliday junction processing in mammalian cells.

During genetic recombination and the recombinational repair of chromosome breaks, DNA molecules become linked at points of strand exchange. Branch migration and resolution of these crossovers, or Holliday junctions (HJs), complete the recombination process. Here, we show that extracts from cells carrying mutations in the recombination/repair genes RAD51C or XRCC3 have reduced levels of HJ resolvase activity. Moreover, depletion of RAD51C from fractionated human extracts caused a loss of branch migration and resolution activity, but these functions were restored by complementation with a variety of RAD51 paralog complexes containing RAD51C. We conclude that the RAD51 paralogs are involved in HJ processing in human cells.