ABSTRACT
Six 2,4,6,8-tetrarylbenzo[1,2- d:4,5- d']bisoxazoles (BBOs) were synthesized: three bearing phenyl substituents at the 2- and 6-positions and three bearing perfluorophenyl groups at those positions. The influence of perfluoro-aryl group substitution on the physical, optical, and electronic properties of 2,4,6,8-tetrarylbenzo[1,2- d:4,5- d']bisoxazoles (BBO) was evaluated using both experimental and theoretical methods. The density functional theory (DFT) model was found to be well-matched to the experimental optical data, as evidenced by the UV-vis spectra. Both cyclic voltammetry (CV) and ultraviolet photoelectron spectroscopy (UPS) were used to determine the position of the HOMO with varying results. The values obtained by CV were deeper than those obtained via UPS and correlated well with the theoretical calculations. However, the UPS values were more consistent with the expected outcomes for a system with segregated frontier molecular orbitals (FMOs). The UPS results are also supported by the electrostatic potential maps, which indicate that the electron density within the LUMO and HOMO is nearly completely localized along the 2,6- or 4,8-axis, respectively. The summation of the results indicates that strongly electron-withdrawing groups can be used to selectively tune the LUMO level with minor perturbation of the HOMO, something that is challenging to accomplish in typical donor-acceptor systems.
ABSTRACT
In Saccharomyces cerevisiae, non-coding RNAs, including cryptic unstable transcripts (CUTs), are subject to degradation by the exosome. The Trf4/5-Air1/2-Mtr4 polyadenylation (TRAMP) complex in S. cerevisiae is a nuclear exosome cofactor that recruits the exosome to degrade RNAs. Trf4/5 are poly(A) polymerases, Mtr4 is an RNA helicase, and Air1/2 are putative RNA-binding proteins that contain five CCHC zinc knuckles (ZnKs). One central question is how the TRAMP complex, especially the Air1/2 protein, recognizes its RNA substrates. To characterize the function of the Air1/2 protein, we used random mutagenesis of the AIR1/2 gene to identify residues critical for Air protein function. We identified air1-C178R and air2-C167R alleles encoding air1/2 mutant proteins with a substitution in the second cysteine of ZnK5. Mutagenesis of the second cysteine in AIR1/2 ZnK1-5 reveals that Air1/2 ZnK4 and -5 are critical for Air protein function in vivo. In addition, we find that the level of CUT, NEL025c, in air1 ZnK1-5 mutants is stabilized, particularly in air1 ZnK4, suggesting a role for Air1 ZnK4 in the degradation of CUTs. We also find that Air1/2 ZnK4 and -5 are critical for Trf4 interaction and that the Air1-Trf4 interaction and Air1 level are critical for TRAMP complex integrity. We identify a conserved IWRXY motif in the Air1 ZnK4-5 linker that is important for Trf4 interaction. We also find that hZCCHC7, a putative human orthologue of Air1 that contains the IWRXY motif, localizes to the nucleolus in human cells and interacts with both mammalian Trf4 orthologues, PAPD5 and PAPD7 (PAP-associated domain containing 5 and 7), suggesting that hZCCHC7 is the Air component of a human TRAMP complex.
Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , DEAD-box RNA Helicases/metabolism , DNA-Directed DNA Polymerase/metabolism , DNA-Directed RNA Polymerases/metabolism , Multiprotein Complexes/metabolism , RNA Stability/physiology , RNA, Fungal/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/metabolism , Adaptor Proteins, Signal Transducing/genetics , Amino Acid Motifs , Amino Acid Substitution , Cell Nucleolus/genetics , Cell Nucleolus/metabolism , Chromosomal Proteins, Non-Histone/genetics , Chromosomal Proteins, Non-Histone/metabolism , DEAD-box RNA Helicases/genetics , DNA-Directed DNA Polymerase/genetics , DNA-Directed RNA Polymerases/genetics , Humans , Multiprotein Complexes/genetics , Mutagenesis , Mutation, Missense , RNA Nucleotidyltransferases/genetics , RNA Nucleotidyltransferases/metabolism , RNA, Fungal/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae Proteins/genetics , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
Elaborar un diagnostico de la situación de los planes de preparativos y respuesta del sector salud frente al cambio climatico en los paises climáticos
