The data point to a novel property of Ddc1, modulating the turnover of DNA binding proteins such as RPAp70 by the proteasome.ĭdc1 checkpoint protein Double-stranded/single-stranded DNA junctions Photoaffinity labeling Proteasome Protein degradation Replication protein A (RPA).Ĭopyright © 2014 Elsevier B.V. The results show that RPAp70 is also subject to proteolysis without photocrosslinking to DNA upon incubation in ddc1Δ extract. We also addressed the question of the stability of free RPA, using anti-RPA antibodies. The degradation of the RPAp70-DNA adduct in ddc1Δ extract is strongly reduced in the presence of the proteasome inhibitor MG 132. Every Gram is hand packaged and inspected to ensure the utmost quality. Providing a top quality clean product with naturally preserved terpenes ensureing you get a smooth flavorful product. The results show that RPAp70 crosslinked to DNA with a 5'-junction is subject to limited proteolysis in ddc1Δ extracts, whereas it is stable in WT, rad17Δ, mec3Δ and mec1Δ extracts. Dabble Extracts is an Award-Winning extraction company boasting 0-PPM on all our product. Interestingly, ddc1Δ extracts did not display photocrosslinking of RPAp70 at a 5'-junction. On the other hand, RPAp70, the large subunit of the replication protein A (RPA), was the predominant crosslinking product at a 5'-junction. In wild-type extract, one of the components of the Ddc1-Rad17-Mec3 complex, Ddc1, was found to be preferentially photocrosslinked at a 3'-junction. Identification of labeled proteins was achieved by MALDI-TOF mass spectrometry peptide mass fingerprinting and genetic analysis. To characterize proteins that interact with single-stranded/double-stranded (ss/ds) DNA junctions in whole cell free extracts of Saccharomyces cerevisiae, we used -labeled photoreactive partial DNA duplexes containing a 3'-ss/ds-junction (3'-junction) or a 5'-ss/ds-junction (5'-junction).
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |