The heterodimer Ku70/80 Ku is the DNA-binding component of the DNA-PK complex required for the nonhomologous end-joining pathway. It participates in numerous nuclear processes, including telomere and chromatin structure maintenance, replication, and transcription. Ku interacts with retroviral preintegration complexes and is thought to interfere with the retroviral replication cycle, in particular the formation of 2-long terminal repeat (LTR) viral DNA circles, viral DNA integration, and transcription. We describe here the effect of Ku80 on both provirus integration and the resulting transgene expression in cells transduced with retroviral vectors. We found that transgene expression was systematically higher in Ku80-deficient xrs6 cells than in Ku80-expressing CHO cells. This higher expression was observed irrespective of the presence of the viral LTR and was also not related to the nature of the promoter. Real-time PCR monitoring of the early viral replicative steps demonstrated that the absence of Ku80 does not affect the efficiency of transduction. We analyzed the transgene distributions localization in nucleus by applying a three-dimensional reconstruction model to two-dimensional fluorescence in situ hybridization images. This indicated that the presence of Ku80 resulted in a bias toward the transgenes being located at the periphery of the nucleus associated with their being repressed; in the absence of this factor the transgenes tend to be randomly distributed and actively expressed. Therefore, although not strictly required for retroviral integration, Ku may be involved in targeting retroviral elements to chromatin domains prone to gene silencing.

The estimation of the fractal dimension in the case of concave log-log Richardson-Mandelbrot plots can be obtained by using asymptotic fractal equations. We demonstrate here, under asymptotic fractal conditions, that additional derivations making use of the Minkowski dilation in grey-scales lead to two asymptotes, one having a slope of 1 and the other a slope of DT-D + 1 (where DT is the topological dimension and D the fractal dimension). The resulting equation offers important advantages. It allows: (i) evaluation of scaling properties of a grey-scale image; (ii) estimation of D without any iteration and (iii) generation of texture and heterogeneity models. We concentrate here on the first two possibilities. Images from cultured cells in studies of cytoskeleton intermediate filaments and kinetic deformability of endothelial cells were used as examples.

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