A5102792600- RNA and protein synthesis mechanisms
- RNA modifications and cancer
- CRISPR and Genetic Engineering
- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- Bacterial Genetics and Biotechnology
- Retinal Development and Disorders
- Neurobiology and Insect Physiology Research
- Photoreceptor and optogenetics research
- Pancreatic function and diabetes
- Caveolin-1 and cellular processes
- Adenosine and Purinergic Signaling
- Enzyme Structure and Function
- Erythrocyte Function and Pathophysiology
- Bacteriophages and microbial interactions
- Stress Responses and Cortisol
- Ion channel regulation and function
- Peptidase Inhibition and Analysis
- Chemical Synthesis and Analysis
- RNA Research and Splicing
- Neuroscience and Neural Engineering
- Cellular transport and secretion
- Sulfur Compounds in Biology
- Cancer-related Molecular Pathways
- Innovation and Socioeconomic Development
University of Colorado Boulder
2003-2022
Horizon Discovery Group (United States)
2018-2022
Institute of Protein Research
2021-2022
Petersburg Nuclear Physics Institute
2018-2021
Peter the Great St. Petersburg Polytechnic University
2018-2019
Kurchatov Institute
2018
Thermo Fisher Scientific (United States)
2008-2015
New York University
2007
Cornell University
2005
Bayer (Italy)
2001
We have shown that ouabain activates Src, resulting in subsequent tyrosine phosphorylation of multiple effectors. Here, we tested if the Na + /K -ATPase and Src can form a functional signaling complex. In LLC-PK1 cells colocalized plasma membrane. Fluorescence resonance energy transfer analysis indicated both proteins were close proximity, suggesting direct interaction. GST pulldown assay showed direct, ouabain-regulated, multifocal interaction between α1 subunit Src. Although kinase domain...
We report the identification of βIV spectrin, a novel spectrin isolated as an interactor receptor tyrosine phosphatase-like protein ICA512. The gene is located on human and mouse chromosomes 19q13.13 7b2, respectively. Alternative splicing generates at least four distinct isoforms, numbered βIVΣ1–βIVΣ4 spectrin. longest isoform (βIVΣ1 spectrin) includes actin-binding domain, followed by 17 repeats, specific domain in which amino acid sequence ERQES repeated times, several putative...
Pairing between the hexamer seed region of a small interfering RNA (siRNA) guide strand (nucleotides 2–7) and complementary sequences in 3′ UTR mature transcripts has been implicated as an important element off-target gene regulation false positive phenotypes. To better understand association profiles we performed analysis all possible (4096) hexamers identified nonuniform distribution frequencies across transcriptome. Subsequent microarray cells transfected with siRNAs having seeds low,...
It has become clear that spinal cord glia (microglia and astrocytes) importantly contribute to the creation of exaggerated pain responses. One model used study this is peri-spinal (intrathecal, i.t.) administration gp120, an envelope protein HIV-1 known activate glia. Previous studies demonstrated i.t. gp120 produces facilitation via release glial proinflammatory cytokines. The present series tested whether nitric oxide (NO) contributes gp120-induced mechanical allodynia and, if so, what...
During amino acid starvation the Escherichia coli stringent response factor RelA recognizes deacylated tRNA in ribosomal A-site. This interaction activates RelA-mediated synthesis of alarmone nucleotides pppGpp and ppGpp, collectively referred to as (p)ppGpp. These two alarmones are synthesized by addition a pyrophosphate moiety 3' position abundant cellular nucleotide GTP less GDP, respectively. Using untagged native we show that allosteric activation increases efficiency GDP conversion...
During host colonization, bacteria use the alarmones (p)ppGpp to reshape their proteome by acting pleiotropically on DNA, RNA, and protein synthesis. Here, we elucidate how initiating ribosome senses cellular pool of guanosine nucleotides regulates progression towards Our results show that affinity triphosphate (GTP) inhibitory concentration ppGpp for 30S-bound initiation factor IF2 vary depending programmed mRNA. The TufA mRNA enhanced GTP 30S complexes, resulting in improved tolerance...
MAD7 is an engineered class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) system isolated from Eubacterium rectale. Analogous to Cas9, it RNA-guided nuclease with demonstrated gene editing activity in Escherichia coli and yeast cells. Here, we report that capable of generating indels fluorescent tagging endogenous genes human HCT116 U2OS cancer cell lines, respectively. In addition, highly proficient indels, small DNA insertions (23 bases), larger integrations ranging 1 14 kb size mouse rat embryos,...
The CRISPR-Cas9 system has been utilized for large-scale, loss-of-function screens mainly using lentiviral pooled formats and cell-survival phenotypic assays. Screening in an arrayed format expands the types of readouts that can be used to now include high-content, morphology-based assays, with recent availability synthetic crRNA libraries, new studies are emerging. Here, we use a cell cycle reporter line perform arrayed, crRNA:tracrRNA screen targeting 169 genes (>600 crRNAs) high content...
Ribosome biogenesis is a highly coordinated and complex process that requires numerous assembly factors ensure prompt flawless maturation of ribosomal subunits. Despite the increasing amount data collected, exact role most mechanistic details their operation remain unclear, mainly due to shortage high-resolution structural information. Here, using cryo-electron microscopy, we characterized 30S particles isolated from an Escherichia coli strain with deleted gene for RbfA factor. The cryo-EM...
Amicoumacin A (Ami) halts bacterial growth by inhibiting the ribosome during translation. The Ami binding site locates in vicinity of E-site codon mRNA. However, does not clash with mRNA, rather stabilizes it, which is relatively unusual and implies a unique way translation inhibition. In this work, we performed kinetic thermodynamic investigation influence on main steps polypeptide synthesis. We show that reduces rate functional canonical 70S initiation complex (IC) formation 30-fold....
Paraneoplastic neurological disorders may result from autoimmunity directed against antigens shared by the affected neurons and associated cancer cells. We have recently reported case of a woman with breast paraneoplastic lower motor neuron syndrome whose serum contained autoantibodies axon initial segments nodes Ranvier myelinated axons, including axons motoneurons. Here, we show that major targets this patient are βIVΣ1 spectrin βIV 140, two isoforms novel gene, as well neuronal surface...
We have previously reported that inescapable tail shock (IS) produces persistent changes in hypothalamic-pituitary-adrenal (HPA) axis function. These are manifest as an elevation basal corticosterone (CORT) levels, a sensitization of adrenocorticotropin hormone (ACTH) and CORT responses to subsequent challenge, failure dexamethasone suppress both the ACTH challenge. The experiments presented here examine IS-induced alterations responsiveness HPA axis, particularly at level anterior...
CHO cells expressing the human insulin receptors (IR) were used to evaluate effect of potent farnesyltransferase inhibitor, manumycin, on antiapoptotic function. Cell treatment with manumycin blocked insulin's ability suppress pro-apoptotic caspase-3 activity which led time-dependent proteolytic cleavage two nuclear target proteins. The Raf-1/MEK/ERK cascade and serine/threonine protein kinase Akt are survival pathways that may be activated in response insulin. We tested hypothesis...
In this study, we used maleimidobutyrylbiocytin to examine possible alteration that may occur in the redox state of insulin receptor (IR) sulfhydryl groups response reduced glutathione (GSH) or N-acetyl-l-cysteine (NAC). Short-term treatment intact cells expressing large numbers IR with GSH NAC led a rapid and reversible reduction α-subunit disulfides, without affecting β-subunit thiol reactivity. The overall integrity oligomeric structure was maintained, indicating neither class I nor II...
While CRISPR interference (CRISPRi) systems have been widely implemented in pooled lentiviral screening, there has limited use with synthetic guide RNAs for the complex phenotypic readouts enabled by experiments arrayed format. Here we describe a novel deactivated Cas9 fusion protein, dCas9-SALL1-SDS3, which produces greater target gene repression than first or second generation CRISPRi when used chemically modified single (sgRNAs), while exhibiting high specificity. We show that...
The growth of the polypeptide chain occurs due to fast and coordinated work ribosome protein elongation factors, EF-Tu EF-G. However, exact contribution each these components in overall balance translation kinetics remains not fully understood. We created an vitro system Escherichia coli replacing either factor with heterologous thermophilic from Thermus thermophilus. rates A-site binding decoding reactions decreased order magnitude presence EF-Tu, indicating that aminoacyl-tRNA delivery...