The dependence of protein splicing on conserved residues of the Cne PRP8 intein was assessed by alanine scanning mutagenesis in a foreign protein context. Corroboration was obtained for the involvement of residues at the splice junctions and of the conserved threonine and histidine of motif B. Five additional residues were identified as absolutely required for splicing. Variant W151A displayed premature C-terminal cleavage, not seen with other Cne PRP8 mutants. We propose a model whereby W151 acts to prevent premature C-terminal cleavage, favoring complete splicing as opposed to two disjointed cleavage events.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.febslet.2007.05.060 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Effect of insertion of intein to Cryptococcus amylolentus, a nonpathogenic fungus closely related to causative agents of cryptococcosis.
Microb Pathog
December 2024
Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, 1703 E Mabel St, Tucson AZ, 85721-0207, USA; The BIO5 Institute, The University of Arizona, Tucson, AZ 85721, USA; Biological Chemistry Program, Department of Chemistry and Biochemistry, College of Science & College of Medicine, The University of Arizona, Tucson, AZ 85721, USA; Department of Molecular & Cellular Biology, College of Science, The University of Arizona, Tucson, AZ 85721, USA. Electronic address:
Inteins are mobile elements within a host protein, with flanking exteins. Autocleavage of intein results in the fusion of exteins, leading to activation of protein. The presence of intein is species dependent.
View Article and Find Full Text PDFEfficient splicing of the CPE intein derived from directed evolution of the PRP8 intein.
Acta Biochim Biophys Sin (Shanghai)
July 2023
College of Biological Science and Medical Engineering, Donghua University, Shanghai 201620, China.
Intein-mediated protein splicing has been widely used in protein engineering; however, the splicing efficiency and extein specificity usually limit its further application. Thus, there is a demand for more general inteins that can overcome these limitations. Here, we study the -splicing of CPE intein obtained from the directed evolution of PRP8, which shows that its splicing rate is ~29- higher than that of the wild-type.
View Article and Find Full Text PDFCalcimycin Inhibits and by Targeting the Prp8 Intein Splicing.
ACS Infect Dis
September 2022
Department of Pharmacology and Toxicology, College of Pharmacy, The University of Arizona, Tucson Arizona 85721-0207, United States.
Drug resistance is a significant concern in the treatment of diseases, including cryptococcosis caused by () and (). Alternative drug targets are necessary to overcome drug resistance before it attains a critical stage. Splicing of inteins from pro-protein precursors is crucial for activities of essential proteins hosting intein elements in many organisms, including human pathogens such as and .
View Article and Find Full Text PDFSmall-molecule inhibitors for the Prp8 intein as antifungal agents.
Proc Natl Acad Sci U S A
January 2021
Wadsworth Center, New York State Department of Health, Albany, NY 12208;
Self-splicing proteins, called inteins, are present in many human pathogens, including the emerging fungal threats () and (), the causative agents of cryptococcosis. Inhibition of protein splicing in sp. interferes with activity of the only intein-containing protein, Prp8, an essential intron splicing factor.
View Article and Find Full Text PDFSpliceosomal Prp8 intein at the crossroads of protein and RNA splicing.
PLoS Biol
October 2019
Department of Biological Sciences and RNA Institute, University at Albany, Albany, New York, United States of America.
The spliceosome is a large ribonucleoprotein complex that removes introns from pre-mRNAs. At its functional core lies the essential pre-mRNA processing factor 8 (Prp8) protein. Across diverse eukaryotes, this protein cofactor of RNA catalysis harbors a self-splicing element called an intein.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!