A5050798487- Peptidase Inhibition and Analysis
- Ubiquitin and proteasome pathways
- RNA Research and Splicing
- Genomics and Chromatin Dynamics
- PARP inhibition in cancer therapy
- Chemical Synthesis and Analysis
- DNA Repair Mechanisms
- Cancer-related gene regulation
- Nuclear Structure and Function
- Machine Learning in Bioinformatics
- Integrated Circuits and Semiconductor Failure Analysis
- Advanced biosensing and bioanalysis techniques
- RNA and protein synthesis mechanisms
- RNA regulation and disease
- RNA modifications and cancer
- Skin and Cellular Biology Research
- Protein Degradation and Inhibitors
- Protein Structure and Dynamics
- Amino Acid Enzymes and Metabolism
- Toxin Mechanisms and Immunotoxins
- Histone Deacetylase Inhibitors Research
- Electrostatic Discharge in Electronics
- HIV Research and Treatment
- Bone and Dental Protein Studies
- Boron Compounds in Chemistry
The University of Texas Southwestern Medical Center
2018-2025
Southwestern Medical Center
2018-2024
Princeton University
2016-2019
University of Pennsylvania
2011-2016
Cancer Research Institute
2014-2016
The Wistar Institute
2010-2014
The X-linked lethal Ogden syndrome was the first reported human genetic disorder associated with a mutation in an N-terminal acetyltransferase (NAT) gene.The affected males harbor Ser37Pro (S37P) gene encoding Naa10, catalytic subunit of NatA, major NAT involved co-translational acetylation proteins.Structural models and molecular dynamics simulations NatA its S37P mutant highlight differences regions catalysis at interface between Naa10 auxiliary hNaa15.Biochemical data further demonstrate...
Protein domains of low sequence complexity do not fold into stable, three-dimensional structures. Nevertheless, proteins with these sequences assist in many aspects cell organization, including assembly nuclear and cytoplasmic structures surrounded by membranes. The dynamic nature cellular assemblies is caused the ability low-complexity (LCDs) to transiently self-associate through labile, cross-β Mechanistic studies useful for study LCD self-association have evolved over past decade form...
Low complexity (LC) head domains 92 and 108 residues in length are, respectively, required for assembly of neurofilament light (NFL) desmin intermediate filaments (IFs). As studied isolation, these IF interconvert between states conformational disorder labile, β-strand-enriched polymers. Solid-state NMR (ss-NMR) spectroscopic studies NFL domain polymers reveal spectral patterns consistent with structural order. A combination intein chemistry segmental isotope labeling allowed preparation...
We report here chemoenzymatic and fully synthetic methodologies to modify aspartate glutamate side chains with ADP-ribose at specific sites on peptides. Structural analysis of ADP-ribosylated peptides reveals near-quantitative migration the chain linkage from anomeric carbon 2″- or 3″-ADP-ribose hydroxyl moieties. find that this pattern is unique ADP-ribosylation propose observed isomer distribution profile present in biochemical cellular environments. After defining distinct stability...
The co-translational modification of N-terminal acetylation is ubiquitous among eukaryotes and has been reported to have a wide range biological effects. human acetyltransferase (NAT) Naa50p (NAT5/SAN) acetylates the α-amino group proteins containing an methionine residue essential for proper sister chromatid cohesion chromosome condensation. elevated activity NATs also correlated with cancer, making these enzymes attractive therapeutic targets. We report x-ray crystal structure bound native...
Abstract The chromatin remodeler ALC1 is activated by DNA damage-induced poly(ADP-ribose) deposited PARP1/PARP2 and their co-factor HPF1. has emerged as a cancer drug target, but how it recruited to ADP-ribosylated nucleosomes affect positioning near breaks unknown. Here we find that PARP1/HPF1 preferentially initiates ADP-ribosylation on the histone H2B tail closest break. To dissect consequences of such asymmetry, generate with defined one side only. cryo-electron microscopy structure...
Abstract Protein domains of low sequence complexity are unable to fold into stable, three-dimensional structures. In test tube studies, these unusual polypeptide regions can self-associate in a manner causing phase separation from aqueous solution. This form protein:protein interaction has been implicated numerous examples dynamic morphological organization within eukaryotic cells. several cases, the basis for domain (LCD) self-association and traced formation labile cross-β The primary...
Significance There are six human amino-terminal acetyltransferases (NATs) that mediate a growing number of biological processes and differentiated from one another on the basis substrate specificity. Interestingly, only more promiscuous NAT enzyme exists in archaea. The crystal structure an archaeal Sulfolobus solfataricus (ssNAT), together with mutagenesis kinetic analysis, reveal active site ssNAT represents hybrid known eukaryotic sites. We highlight features this protein allow it to...
Significance Programmable DNA-binding proteins such as nuclease-deficient Cas9 (dCas9) offer a simple system for genomic localization of genetically encodable biomolecules. These tools have enabled characterization numerous different chromatin effectors at specific genetic elements. The delivery fully synthetic cargos to loci is currently limited by lack adequate technologies. Here we used protein trans -splicing ligate chemical moieties dCas9 in vitro and subsequently deliver these...
Serine ADP-ribosylation (ADPr) is a DNA damage-induced post-translational modification catalyzed by the PARP1/2:HPF1 complex. As list of substrates continues to expand, there need for technologies prepare mono- and poly-ADP-ribosylated proteins biochemical interrogation. Here, we investigate unique peptide ADPr activities PARP1 in absence presence HPF1. We then exploit these develop method that facilitates installation ADP-ribose polymers onto peptides with precise control over chain length...
Protein acetylation is a prevalent posttranslational modification that regulated by diverse acetyltransferase enzymes. Although histone acetyltransferases (HATs) have been well characterized both structurally and mechanistically, far less known about non-histone The human ESCO1 ESCO2 paralogs acetylate the cohesin complex subunit SMC3 to regulate separation of sister chromatids during mitosis meiosis. Missense mutations within domain these proteins correlate with diseases, including...
Recently developed chemical and enzyme-based technologies to install serine ADP-ribosylation onto synthetic peptides have enabled new approaches study poly(ADP-ribose) polymerase (PARP) biology. Here, we establish a generalizable strategy prepare ADP-ribosylated that are compatible with N-terminal, C-terminal, sequential protein ligation reactions. Two unique protein-assembly routes employed generate full-length linker histone constructs homogeneously at known DNA damage-dependent...
Summary Disease-causing missense mutations that occur within structurally and functionally unannotated protein regions can guide researchers to new mechanisms of regulation dysfunction. Here, we report the thrombocytopenia-, myelodysplastic syndromes-, leukemia-associated P214L mutation in transcriptional regulator ETV6 creates an XPO1-dependent nuclear export signal cause mislocalization. Strategies disrupt XPO1 activity fully restore localization transcription activity. Mechanistic insight...
Nt-acetylation is among the most common protein modifications in eukaryotes. Although thought for a long time to protect proteins from degradation, role of still debated. It catalyzed by enzymes called N-terminal acetyltransferases (NATs). In eukaryotes, several NATs, composed at least one catalytic domain, target different substrates based on their sequences. order better understand substrate specificity human we investigated silico enzyme-substrate interactions four subunits NATs (Naa10p,...
Germline and somatic missense mutations in the transcriptional repressor ETV6 are strongly associated with blood diseases, including thrombocytopenia leukemia. These dominant negative mutations, which scattered throughout DNA binding domain a functionally unannotated linker region, cause protein to mislocalize cytoplasm. We set out answer following question: why do seemingly unrelated similar mislocalization phenotypes? A combination of chemical, biochemical, cellular approaches revealed...
Certain protein intrinsically disordered regions (IDRs) reversibly form higher-order oligomers to fulfill their cellular functions. As IDRs lack predictable structured elements, determination of sequence features that encode IDR oligomerization is an active area study. In particular, the contribution transiently-structured interactions remains challenging assess using conventional biochemical and biophysical techniques. To surmount this challenge, a recent study from our group utilized...