A5083677717- RNA Research and Splicing
- RNA modifications and cancer
- Neurogenetic and Muscular Disorders Research
- Genomics and Chromatin Dynamics
- RNA and protein synthesis mechanisms
- Epigenetics and DNA Methylation
- Cancer-related gene regulation
- Chromosomal and Genetic Variations
- Nuclear Structure and Function
- CRISPR and Genetic Engineering
- Circular RNAs in diseases
- RNA regulation and disease
- Cancer-related molecular mechanisms research
- Congenital Anomalies and Fetal Surgery
- Microtubule and mitosis dynamics
- Genomics and Phylogenetic Studies
- DNA Repair Mechanisms
- Molecular Biology Techniques and Applications
- Genomic variations and chromosomal abnormalities
- Advanced biosensing and bioanalysis techniques
- Animal Genetics and Reproduction
- Cardiomyopathy and Myosin Studies
- Cancer-related Molecular Pathways
- Peptidase Inhibition and Analysis
- Ubiquitin and proteasome pathways
University of North Carolina at Chapel Hill
2016-2025
UNC Lineberger Comprehensive Cancer Center
2015-2024
Segeberger Kliniken
2016-2018
Pediatrics and Genetics
2018
Case Western Reserve University
2000-2009
University School
1997-2007
University Hospitals of Cleveland
1996-2002
Center for Human Genetics
1994-2002
Massachusetts General Hospital
2000
Brigham and Women's Hospital
2000
The D-type cyclin-dependent kinases CDK4 and CDK6 are complexed with many small cellular proteins (p14, p15, p16, p18, p20). We have isolated cDNA sequences corresponding to the MTS2 genomic fragment that encodes CDK4- CDK6-associated p14 protein. By use of a yeast interaction screen search for CDK6-interacting proteins, we also identified an 18-kD human protein, is homolog cyclin D-CDK4 inhibitors p16 (INK4A/MTS1) (MTS2/INK4B). Both in vivo vitro, p18 interacts strongly CDK6, weakly CDK4,...
In eukaryotic cells, histone gene expression is one of the major events that mark entry into S phase. While this process tightly linked to cell cycle position, how it regulated by machinery not known. Here we show NPAT, a substrate cyclin E–Cdk2 complex, associated with human replication-dependent clusters on both chromosomes 1 and 6 in We demonstrate NPAT activates transcription activation dependent promoter elements (SSCSs) previously proposed mediate cycle–dependent transcription. Cyclin...
Although bulk chromatin is thought to have limited mobility within the interphase eukaryotic nucleus, directed long-distance chromosome movements are not unknown. Cajal bodies (CBs) nuclear suborganelles that nonrandomly associate with small RNA (snRNA) and histone gene loci in human cells during interphase. However, mechanism responsible for this association uncertain. In study, we present an experimental system probe dynamic interplay of CBs a U2 snRNA target locus transcriptional...
We report the discovery of a class abundant circular noncoding RNAs that are produced during metazoan tRNA splicing. These transcripts, termed intronic (tric)RNAs, conserved features animal transcriptomes. Biogenesis tricRNAs requires anciently sequence motifs and processing enzymes, their expression is regulated in an age-dependent tissue-specific manner. Furthermore, we exploited this biogenesis pathway to develop vivo system for generating “designer” human cells. Reporter constructs...
Coiled bodies (CBs) are nuclear organelles whose structures appear to be highly conserved in evolution. In rapidly cycling cells, they typically located the nucleoplasm but often found contact with nucleolus. The CBs human cells contain a unique protein, called p80-coilin. Studies on amphibian oocyte nuclei have revealed protein within "sphere" organelle that shares significant structural similarity Spheres and also enriched small ribonucleoproteins other RNA-processing components. We...
Spinal muscular atrophy (SMA) is a genetic disorder caused by mutations in the human survival of motor neuron 1 gene, SMN1 . SMN protein part large complex that required for biogenesis various small nuclear ribonucleoproteins (snRNPs). Here, we report interacts directly with Cajal body signature protein, coilin, and this interaction mediates recruitment to bodies. Mutation or deletion specific RG dipeptide residues within coilin inhibits both vivo vitro. Interestingly, GST-pulldown...
Cajal bodies (CBs) are nuclear suborganelles involved in the biogenesis of small ribonucleoproteins (snRNPs). In addition to snRNPs, they highly enriched basal transcription and cell cycle factors, nucleolar proteins fibrillarin (Fb) Nopp140 (Nopp), survival motor neuron (SMN) protein complex, CB marker protein, p80 coilin. We report generation knockout mice lacking COOH-terminal 487 amino acids Northern Western blot analyses demonstrate that we have successfully removed full-length coilin...
Cajal bodies (CBs) are subnuclear domains implicated in small nuclear ribonucleoprotein (snRNP) biogenesis. In most cell types, CBs coincide with gems, which contain the survival of motor neurons (SMN) complex, an essential snRNP assembly factor. Here, we analyze exchange kinetics multiple components and gems living cells using photobleaching microscopy. We demonstrate differences dissociation CB constituents relate them to their functions. Coilin SMN complex members exhibit relatively long...
Spinal muscular atrophy (SMA) is a neurodegenerative disease characterized by loss of spinal motor neurons. The gene encoding the survival neurons (SMN) protein mutated in >95% SMA cases. SMN central component large oligomeric complex, including Gemins2-7, that necessary and sufficient for vivo assembly Sm proteins onto small nuclear (sn)RNAs mediate pre-mRNA splicing. After cytoplasmic core, both splicing snRNPs are imported into nucleus, accumulating Cajal bodies additional snRNA...
Mutations in human survival motor neurons 1 (SMN1) cause spinal muscular atrophy (SMA) and are associated with defects assembly of small nuclear ribonucleoproteins (snRNPs) vitro. However, the etiological link between snRNPs SMA is unclear. We have developed a Drosophila melanogaster system to model vivo. Larval-lethal Smn-null mutations show no detectable snRNP reduction, making it unlikely that these animals die from global deprivation. Hypomorphic Smn reduce dSMN protein levels adult...
Cajal bodies (CBs) are nuclear organelles that occur in a variety of organisms, including vertebrates, insects, and plants. They most often identified with antibodies against the marker protein coilin. Because amino acid sequence coilin is not strongly conserved evolutionarily, orthologues have been difficult to recognize by homology search. Here, we report identification Drosophila melanogaster describe its distribution tissues fly. Surprisingly, found only CBs but also histone locus...
Mature tRNAs are generated by multiple post-transcriptional processing steps, which can include intron removal. Recently, we discovered a new class of circular non-coding RNAs in metazoans, called tRNA intronic (tric)RNAs. To investigate the mechanism tricRNA biogenesis, constructs that replace native introns human and fruit fly genes with Broccoli fluorescent RNA aptamer. Using these reporters, identified cis-acting elements required for formation vivo. Disrupting conserved base pair...
Circular (circ)RNAs have recently become a subject of great biologic interest. It is now clear that they represent diverse and abundant class RNAs with regulated expression evolutionarily conserved functions. There are several mechanisms by which RNA circularization can occur in vivo. Here, we focus on the biogenesis tRNA intronic circular (tricRNAs) archaea animals, detail their use as research tools for orthogonal, directed circRNA
We have found that coilin, the marker protein for Cajal bodies (coiled bodies, CBs), is a self-interacting protein, and we mapped domain responsible this activity to amino-terminus. Together with nuclear localization signal, self-interaction necessary sufficient CBs. Overexpression of various wild-type mutant coilin constructs in HeLa cells results disruption both CBs survival motor neurons (SMN) gems. Additionally, identified cryptic nucleolar signal (NoLS), within which may be exposed...
The human parvovirus adeno-associated virus (AAV) is unique in its ability to target viral integration a specific site on chromosome 19 (ch-19). Recombinant AAV (rAAV) vectors retain the integrate but have apparently lost this target. In report, we characterize terminal-repeat-mediated for wild-type (wt), rAAV, and vitro systems gain better understanding of these differences. Cell lines latent either wt or rAAV were characterized by variety techniques, including PCR, Southern hybridization,...