ORCID Profile
0000-0001-7683-3293
Current Organisation
University of St Andrews
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Publisher: Springer Science and Business Media LLC
Date: 22-07-2022
DOI: 10.1038/S41596-022-00708-4
Abstract: Multiple aspects of mRNA translation are subject to regulation. Here we present a ribosome footprinting protocol to determine the location and composition of 40S and 80S ribosome complexes on endogenous mRNAs transcriptome-wide in vivo in yeast and mammalian cells. We present an extension of the translation complex profiling (TCP-seq) protocol, originally developed in yeast, by including an immunoprecipitation step to assay the location of both 40S and 80S ribosome complexes containing proteins of interest. This yields information on where along mRNAs the ribosome-bound protein of interest joins the ribosome to act, and where it leaves again, thereby monitoring the sequential steps of translation and the roles of various translation factors therein. Rapid fixation of live cells ensures the integrity of all translation complexes bound to mRNA at native positions. Two procedures are described, differing mainly in the fixation conditions and the library preparation. Depending on the research question, either procedure offers advantages. Execution of a Sel-TCP-seq experiment takes 5-10 working days, and initial data analysis can be completed within 2 days.
Publisher: Elsevier BV
Date: 08-2020
Publisher: Wiley
Date: 04-08-2022
Abstract: Dimagnesium(I) complexes [{( Ar nacnac)Mg} 2 ], where Ar nacnac=HC(MeCNAr) 2 , Ar=Dip=2,6‐ i Pr 2 −C 6 H 3 , Ar=Dep=2,6‐Et 2 −C 6 H 3 , Ar=Mes=2,4,6‐Me 3 −C 6 H 2 , react with iodoarenes in oxidative addition reactions. With iodobenzene, magnesium phenyl and magnesium iodide complex fragments were obtained, and from a reaction with 4‐iodo‐1,2,3,5‐tetramethylpyrazolium iodide, [ Me PZI]I, the pyrazol‐4‐ylidene complex [( Dip nacnac)MgI( Me PZ)] was structurally characterised, alongside other products. The isomeric imidazol‐2‐ylidene complex [( Dip nacnac)MgI( Me NHC)], where Me NHC is 1,3,4,5‐tetramethylimidazol‐2‐ylidene, was prepared and characterised. X‐ray crystal structure determinations and DFT computational studies have been carried out to compare the related complexes. The results show that the Me PZ ligand is higher in energy and more nucleophilic than its more common isomeric Me NHC carbene.
Publisher: Wiley
Date: 11-07-2022
Abstract: The new β‐diketimine i PrDip nacnacH, HC( i PrCNDip) 2 H, Dip=2,6‐ i Pr 2 ‐C 6 H 3 , was converted to the magnesium(I) complex [{( i PrDip nacnac)Mg} 2 ] and reaction with 2‐adamantanone (OAd) afforded the ketone‐1,2‐diide complex [{( i PrDip nacnac)Mg} 2 (μ‐OAd)]. The complex contains the first stable dianion of an aliphatic ketone with an electropositive metal and shows an OAd 2− unit with long C−O bond and pyramidal carbon centre. DFT studies reveal an anionic charge on both neighbouring C and O atoms. Reductions of aliphatic ketones with magnesium(I) complexes show that these likely proceed via highly reactive dianions and afforded a 1 : 1 mixture of an alkoxide and an enolate when an enolisable ketone was used, and rapid CH activations reactions, e.g., of stabilising ligand moieties, when non‐enolisable ketones were employed.
Publisher: American Chemical Society (ACS)
Date: 25-09-2023
Location: United Kingdom of Great Britain and Northern Ireland
No related grants have been discovered for Tanja van Mourik.