Technical Reports

Filter By:

Article Type
  • The authors describe the development of ASPIR-1, a small molecule that specifically inhibits AAA proteins by covalently modifying a cysteine residue introduced by mutagenesis at the AAA ATPase site.

    • Tommaso Cupido
    • Natalie H. Jones
    • Tarun M. Kapoor
    Technical Report
  • CRISPR–Cas9-based method to introduce site-specific S-GlcNAcylation enables dissection of the roles of protein O-GlcNAcylation. S-GlcNAc, a hydrolytically stable structural mimic of O-GlcNAc, is recognized by a range of O-GlcNAc-binding proteins.

    • Andrii Gorelik
    • Sergio Galan Bartual
    • Daan M. F. van Aalten
    Technical Report
  • Applying SHAPE-seq to analyze cotranscriptional folding of the B. cereus crcB fluoride riboswitch at nucleotide resolution shows that the folding pathway undergoes a ligand-dependent bifurcation that influences terminator formation via coordinated structural transitions.

    • Kyle E Watters
    • Eric J Strobel
    • Julius B Lucks
    Technical Report
  • READ is a new crystallographic approach to visualize conformational ensembles of heterogeneous and dynamic molecules. READ is applied here to structurally characterize the various folding states of client Im7 bound to chaperone Spy.

    • Scott Horowitz
    • Loïc Salmon
    • James C A Bardwell
    Technical Report
  • It has been challenging to label endogenous genomic sequences in living cells, and this has limited attempts to study the dynamics of nuclear architecture in genome function. In a newly developed methodology, transcription activator–like effectors (TALEs) were used to label endogenous repetitive genomic sequences to visualize nuclear positioning and chromatin dynamics in cultured mouse cells and embryos.

    • Yusuke Miyanari
    • Céline Ziegler-Birling
    • Maria-Elena Torres-Padilla
    Technical Report
  • SUMOylation is a dynamic protein post-translational modification that regulates many eukaryotic proteins. Now a methodology using commercially available monoclonal antibodies coupled to MS analysis leads to the enrichment and identification of endogenous targets for SUMO1 and for SUMO2/3 in HeLa cells and mouse liver. This protocol can be adapted for other tissues and organs.

    • Janina Becker
    • Sina V Barysch
    • Frauke Melchior
    Technical Report
  • Previous studies have shown the potential for convergent transcription as a way to induce gene silencing. This Technical Report now demonstrates that introducing convergent transcription plasmids into either fission yeast or mammalian cells can be used to mediate long-term transcriptional gene silencing of endogenous genes, with major advantages over other gene silencing strategies.

    • Monika Gullerova
    • Nick J Proudfoot
    Technical Report
  • NMR relaxation phenomena give insight into local protein motions, and understanding protein dynamics can aid in understanding protein function. Nuclear Overhauser enhancements (NOEs) have been traditionally used to determine protein structure by NMR, but a novel application using exact NOEs provides a structural ensemble that describes a protein's structure as well as its dynamics.

    • Beat Vögeli
    • Sina Kazemi
    • Roland Riek
    Technical Report
  • Tryptophan scanning is now used to characterize transient intermediate states in the folding pathway of a protein. By following the intrinsic fluorescence of tryptophan residues introduced at different solvent-exposed positions of model protein ubiquitin, the authors characterize a late folding intermediate.

    • Alexis Vallée-Bélisle
    • Stephen W Michnick
    Technical Report