Expansion of microsatellites on evolutionary young Y chromosome.

Sex chromosomes are an ideal system to study processes connected with suppressed recombination. We found evidence of microsatellite expansion, on the relatively young Y chromosome of the dioecious plant sorrel (Rumex acetosa, XY1Y2 system), but no such expansion on the more ancient Y chromosomes of liverwort (Marchantia polymorpha) and human. The most expanding motifs were AC and AAC, which also showed periodicity of array length, indicating the importance of beginnings and ends of arrays. Our data indicate that abundance of microsatellites in genomes depends on the inherent expansion potential of specific motifs, which could be related to their stability and ability to adopt unusual DNA conformations. We also found that the abundance of microsatellites is higher in the neighborhood of transposable elements (TEs) suggesting that microsatellites are probably targets for TE insertions. This evidence suggests that microsatellite expansion is an early event shaping the Y chromosome where this process is not opposed by recombination, while accumulation of TEs and chromosome shrinkage predominate later.

Interferon-gamma, but not interferon-alpha, induces SOCS 3 expression in human melanoma cell lines.

The signal transducers and transcription activators (STATs) and their endogenous inhibitors of the suppressors of cytokine signalling (SOCS) family are major proteins harmonizing the transmission of external signals from the surface membrane to target genes in the nucleus. To correlate the induction of SOCS 3 by interferons (IFNs) on messenger RNA and protein levels with STAT 1 phosphorylation in human malignant melanoma cell lines, we used a unique collection of 18 established malignant melanoma cell lines and six human non-malignant normal cells (two melanocytes, two skin keratinocytes and two fibroblasts). IFN-gamma induced SOCS 3 in 83% of melanoma cell lines, whereas IFN-alpha stimulated SOCS 3 expression in only 11% of cases. Similarly, melanocytes showed strong induction of SOCS 3 by IFN-gamma and, to a lesser extent, by IFN-alpha. In most cases, SOCS 3 expression was paralleled by STAT 1 phosphorylation at tyrosine residues (Y701). In several lines, however, SOCS 3 was not induced despite STAT 1 phosphorylation and, in a few lines, SOCS 3 induction occurred without detectable STAT 1 phosphorylation, indicating that STAT 1 might not be an exclusive inducer of SOCS 3. Similarly, non-malignant cells displayed STAT 1 activation and high levels of SOCS 3 expression after IFN-gamma (but not IFN-alpha) treatment. In conclusion, in contrast to IFN-alpha, IFN-gamma appeared to induce SOCS 3 apparently at the transcription level and exhibited higher cytotoxic effects regardless of the cell origin.

Malignant melanoma associates with deficient IFN-induced STAT 1 phosphorylation.

STAT 1, a member of signal transducers and transcription activators of STAT family proteins, has been implicated as important mediator of IFN signaling. Functional activation of STAT 1 requires tyrosine and serine phosphorylation. Defects in its expression or activation in response to IFNs were observed in numerous pathological conditions including cancer. To further explore cancer-associated impaired STAT 1 response to IFNs, the inducibility of serine (S 727) and tyrosine (Y 701) phosphorylation by IFN-alpha/-gamma was assessed in 21 melanoma cell lines and in 35 primary cultures derived from melanoma patients. STAT 1 levels and inducibility of its activated phospho-forms were detected by Western analysis using specific polyclonal and monoclonal antibodies. All cell lines as well as patient melanoma samples expressed STAT 1 with variable signal intensity. Significant impaired IFN-induced STAT 1 S 727 phosphorylation was observed in both model systems with average of 77% of non-responders recorded in patient melanoma cells and 76% in melanoma cell lines. Failure of PY 701 induction occurred in patient samples (63% after IFN-alpha and 34% after IFN-gamma induction) and to a lesser degree in cell lines (i.e. response absence to IFN-alpha in 5 and to IFN-gamma in 2 melanoma lines). Our study demonstrates STAT 1 functional abnormalities in melanoma cells. On the basis of detailed analyses of patient melanoma cells with respect to the inducibility of STAT 1 phosphorylation by IFNs, four categories of patients could be distinguished: a) activation on both S 727 and Y 701, b) not inducible response, c) activation on Y 701 but not on S 727, d) heterogeneous response. Clinical study is now in progress to establish the significance of in vitro STAT 1 activation for predicting the response to IFN-based therapy and to explore biological consequences in cases responding in vitro to IFN-induced STAT 1 activation on only one of the critical amino acid residues.