Haloferax volcanii microproteins that contain two CPXCG motifs indicative for one-domain zinc finger proteins

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This project in the framework of the Priority Program SPP2002 “Small proteins in prokaryotes” concentrates on very small proteins of the halophilic archaeaon Haloferax volcanii, which contain two C(P)XCG motifs, and, thus, are one-domain zinc finger proteins. In the first granting period, phenotypic characterization of 16 deletion mutants has revealed the high biological significance of haloarchaeal C(P)XCG proteins. One protein, HVO_2753, has been characterized in detail, and its NMR solution structure was solved.In the second granting period, a deletion mutant collection of further C(P)XCG protein genes will be generated and characterized. A novel approach of “hidden thiol proteomics” will be established and used for the parallel characterization of differential production of small zinc finger proteins under various environmental conditions. HVO_2753 and a few additional C(P)XCG proteins of high biological significance will be further characterized in detail, e.g. the importance of selected single amino acids for protein function in vivo will be unraveled, interaction partners will be identified, and further structural studies will be performed. In summary, the project will lead to an unprecedented insight into the biological roles and molecular mechanisms of small zinc finger proteins in haloarchaea.