73 research outputs found
Tunable Cavity Optomechanics with Ultracold Atoms
We present an atom-chip-based realization of quantum cavity optomechanics
with cold atoms localized within a Fabry-Perot cavity. Effective sub-wavelength
positioning of the atomic ensemble allows for tuning the linear and quadratic
optomechanical coupling parameters, varying the sensitivity to the displacement
and strain of a compressible gaseous cantilever. We observe effects of such
tuning on cavity optical nonlinearity and optomechanical frequency shifts,
providing their first characterization in the quadratic-coupling regime.Comment: 4 pages, 5 figure
DNA Renaturation at the Water-Phenol Interface
We study DNA adsorption and renaturation in a water-phenol two-phase system,
with or without shaking. In very dilute solutions, single-stranded DNA is
adsorbed at the interface in a salt-dependent manner. At high salt
concentrations the adsorption is irreversible. The adsorption of the
single-stranded DNA is specific to phenol and relies on stacking and hydrogen
bonding. We establish the interfacial nature of a DNA renaturation at a high
salt concentration. In the absence of shaking, this reaction involves an
efficient surface diffusion of the single-stranded DNA chains. In the presence
of a vigorous shaking, the bimolecular rate of the reaction exceeds the
Smoluchowski limit for a three-dimensional diffusion-controlled reaction. DNA
renaturation in these conditions is known as the Phenol Emulsion Reassociation
Technique or PERT. Our results establish the interfacial nature of PERT. A
comparison of this interfacial reaction with other approaches shows that PERT
is the most efficient technique and reveals similarities between PERT and the
renaturation performed by single-stranded nucleic acid binding proteins. Our
results lead to a better understanding of the partitioning of nucleic acids in
two-phase systems, and should help design improved extraction procedures for
damaged nucleic acids. We present arguments in favor of a role of phenol and
water-phenol interface in prebiotic chemistry. The most efficient renaturation
reactions (in the presence of condensing agents or with PERT) occur in
heterogeneous systems. This reveals the limitations of homogeneous approaches
to the biochemistry of nucleic acids. We propose a heterogeneous approach to
overcome the limitations of the homogeneous viewpoint
Crystal Structure of TDRD3 and Methyl-Arginine Binding Characterization of TDRD3, SMN and SPF30
SMN (Survival motor neuron protein) was characterized as a dimethyl-arginine binding protein over ten years ago. TDRD3 (Tudor domain-containing protein 3) and SPF30 (Splicing factor 30 kDa) were found to bind to various methyl-arginine proteins including Sm proteins as well later on. Recently, TDRD3 was shown to be a transcriptional coactivator, and its transcriptional activity is dependent on its ability to bind arginine-methylated histone marks. In this study, we systematically characterized the binding specificity and affinity of the Tudor domains of these three proteins quantitatively. Our results show that TDRD3 preferentially recognizes asymmetrical dimethylated arginine mark, and SMN is a very promiscuous effector molecule, which recognizes different arginine containing sequence motifs and preferentially binds symmetrical dimethylated arginine. SPF30 is the weakest methyl-arginine binder, which only binds the GAR motif sequences in our library. In addition, we also reported high-resolution crystal structures of the Tudor domain of TDRD3 in complex with two small molecules, which occupy the aromatic cage of TDRD3
The assembly of a spliceosomal small nuclear ribonucleoprotein particle
The U1, U2, U4, U5 and U6 small nuclear ribonucleoprotein particles (snRNPs) are essential elements of the spliceosome, the enzyme that catalyzes the excision of introns and the ligation of exons to form a mature mRNA. Since their discovery over a quarter century ago, the structure, assembly and function of spliceosomal snRNPs have been extensively studied. Accordingly, the functions of splicing snRNPs and the role of various nuclear organelles, such as Cajal bodies (CBs), in their nuclear maturation phase have already been excellently reviewed elsewhere. The aim of this review is, then, to briefly outline the structure of snRNPs and to synthesize new and exciting developments in the snRNP biogenesis pathways
Molecular Determinants of Survival Motor Neuron (SMN) Protein Cleavage by the Calcium-Activated Protease, Calpain
Spinal muscular atrophy (SMA) is a leading genetic cause of childhood mortality, caused by reduced levels of survival motor neuron (SMN) protein. SMN functions as part of a large complex in the biogenesis of small nuclear ribonucleoproteins (snRNPs). It is not clear if defects in snRNP biogenesis cause SMA or if loss of some tissue-specific function causes disease. We recently demonstrated that the SMN complex localizes to the Z-discs of skeletal and cardiac muscle sarcomeres, and that SMN is a proteolytic target of calpain. Calpains are implicated in muscle and neurodegenerative disorders, although their relationship to SMA is unclear. Using mass spectrometry, we identified two adjacent calpain cleavage sites in SMN, S192 and F193. Deletion of small motifs in the region surrounding these sites inhibited cleavage. Patient-derived SMA mutations within SMN reduced calpain cleavage. SMN(D44V), reported to impair Gemin2 binding and amino-terminal SMN association, drastically inhibited cleavage, suggesting a role for these interactions in regulating calpain cleavage. Deletion of A188, a residue mutated in SMA type I (A188S), abrogated calpain cleavage, highlighting the importance of this region. Conversely, SMA mutations that interfere with self-oligomerization of SMN, Y272C and SMNΔ7, had no effect on cleavage. Removal of the recently-identified SMN degron (Δ268-294) resulted in increased calpain sensitivity, suggesting that the C-terminus of SMN is important in dictating availability of the cleavage site. Investigation into the spatial determinants of SMN cleavage revealed that endogenous calpains can cleave cytosolic, but not nuclear, SMN. Collectively, the results provide insight into a novel aspect of the post-translation regulation of SMN
Identification and phylogenetic analyses of the protein arginine methyltransferase gene family in fish and ascidians
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