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Jennifer Gerke, Özgür Bayram, Gerhard H. Braus The velvet complex coordinates light, fungal development and secondary metabolism in Aspergillus nidulans Differentiation and secondary metabolism are correlated processes in fungi and respond to light. In Aspergillus nidulans, light inhibits sexual reproduction as well as secondary metabolism. We identified the heterotrimeric velvet complex as link between lightresponding developmental regulation and control of secondary metabolism. VeA, which is primarily expressed in the dark, physically interacts with VelB that is expressed during sexual development. VeA bridges VelB to the nuclear master regulator of secondary metabolism LaeA. Deletion of either velB or veA results in defects in both sexual fruiting body formation and production of secondary metabolites. contact: Jennifer Gerke Georg-August-University Göttingen Institute of Microbiology and Genetics jgerke@gwdg.de Grisebachstr. 8 37077 Göttingen (Germany)
Filip Senigl, Jiri Plachy, Jiri Hejnar The CpG island core element protects retroviral vectors from transcriptional silencing Unmethylated CpG islands are known to maintain adjacent promoters transcriptionally active. In the adenosine phosphoribosyltransferase gene-adjacent CpG island, the protection from transcriptional silencing can be attributed to the short CpG-rich core element containing Sp1 binding sites. We report here the insertion of this CpG island core element, IE, into the long terminal repeat of a retroviral vector derived from Rous sarcoma virus, which normally suffers from progressive transcriptional silencing in mammalian cells. IE insertion into specific position between enhancer and promoter sequences led to efficient protection of the integrated vector from silencing and gradual CpG methylation in rodent and human cells. Individual cell clones with IE-modified reporter vectors display high level of reporter expression for a sustained period and without substantial variegation in the cell culture. The presence of Sp1 binding sites is necessary for the protective effect of IE. Modified vectors with IE insertions are also refractory to the transcriptional silencing during integration as evidenced by colonyforming assay. We suggest that our strategy of anti-silencing protection by CpG island core element may prove generally useful in retroviral vectors. contact: Jiri Hejnar Institute of Molecular Genetics hejnar@img.cas.cz Videnska 1083 CZ-14220 Prague 6 (Czechia)
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