3,857 research outputs found
Light Regulation of Protein Dimerization and Kinase Activity in Living Cells Using Photocaged Rapamycin and Engineered FKBP
We developed a new system for light-induced protein dimerization in living cells using a novel photocaged analog of rapamycin (pRap) together with an engineered rapamycin binding domain (iFKBP). Using focal adhesion kinase as a target, we demonstrated successful light-mediated regulation of protein interaction and localization in living cells. Modification of this approach enabled light-triggered activation of a protein kinase and initiation of kinase-induced phenotypic changes in vivo
Restriction enzyme-free mutagenesis via the light regulation of DNA polymerization
The effects of photocaged nucleosides on the DNA polymerization reaction was investigated, finding that most polymerases are unable to recognize and read through the presence of a single caging group on the DNA template. Based on this discovery, a new method of introducing mutations into plasmid DNA via a light-mediated mutagenesis protocol was developed. This methodology is advantageous over several common approaches in that it requires the use of only two polymerase chain reaction primers, and does not require any restriction sites or use of restriction enzymes. Additionally, this approach enables not only site-directed mutations, but also the insertion of DNA strands of any length into plasmids and the deletion of entire genes from plasmids
The Human Mitochondrial tRNAMet: Structure/Function Relationship of a Unique Modification in the Decoding of Unconventional Codons
Human mitochondrial mRNAs utilize the universal AUG and the unconventional isoleucine AUA codons for methionine. In contrast to translation in the cytoplasm, human mitochondria use one tRNA, hmtRNAMetCAU, to read AUG and AUA codons at both the peptidyl- (P-), and aminoacyl-(A-) sites of the ribosome. The hmtRNAMetCAU has a unique post-transcriptional modification, 5-formylcytidine, at the wobble position 34 (f5C34), and a cytidine substituting for the invariant uridine at position 33 of the canonical âU-turnâ in tRNAs. The structure of the tRNA's anticodon stem and loop domain (hmtASLMetCAU), determined by NMR restrained molecular modeling, revealed how the f5C34 modification facilitates the decoding of AUA at the P- and A-sites. The f5C34 defined a reduced conformational space for the nucleoside, in what appears to have restricted the conformational dynamics of the anticodon bases of the modified hmtASLMetCAU. The hmtASLMetCAU exhibited a âC-turnâ conformation that has some characteristics of the U-turn motif. Codon binding studies with both E. coli and bovine mitochondrial ribosomes revealed that the f5C34 facilitates AUA binding in the A-site and suggested that the modification favorably alters the ASL's binding kinetics. Mitochondrial translation by many organisms including humans sometimes initiates with the universal isoleucine codons AUU and AUC. The f5C34 enabled P-site codon binding to these normally isoleucine codons. Thus, the physicochemical properties of this one modification, f5C34, expand codon recognition from the traditional AUG to the non-traditional, synonymous codons AUU and AUC as well as AUA, in the reassignment of universal codons in the mitochondria
Search for Neutral Higgs Bosons of the Minimal Supersymmetric Standard Model in e+e- Interactions at \sqrt{s} = 189 GeV
A search for the lightest neutral scalar and neutral pseudoscalar Higgs
bosons in the Minimal Supersymmetric Standard Model is performed using 176.4
pb^-1 of integrated luminosity collected by L3 at a center-of-mass energy of
189 GeV. No signal is observed, and the data are consistent with the expected
Standard Model background. Lower limits on the masses of the lightest neutral
scalar and pseudoscalar Higgs bosons are given as a function of tan(beta).
Lower mass limits for tan(beta)>1 are set at the 95% confidence level to be m_h
> 77.1 GeV and m_A > 77.1 GeV
Search for Neutral Higgs Bosons of the Minimal Supersymmetric Standard Model in e+e- Interactions at sqrt(s) up to 209 GeV
A search for the lightest neutral CP-even and neutral CP-odd Higgs bosons of
the Minimal Supersymmetric Standard Model is performed using 216.6 pb-1 of data
collected with the L3 detector at LEP at centre-of-mass energies between 203
and 209 GeV. No indication of a signal is found. Including our results from
lower centre-of-mass energies, lower limits on the Higgs boson masses are set
as a function of tan(beta) for several scenarios. For tan(beta) greater than
0.7 they are mh > 84.5 GeV and mA > 86.3 GeV at 95% confidence level
Measurement of Exclusive rho+rho- Production in Mid-Virtuality Two-Photon Interactions and Study of the gamma gamma* -> rho rho Process at LEP
Exclusive rho+rho- production in two-photon collisions between a quasi-real
photon, gamma, and a mid-virtuality photon, gamma*, is studied with data
collected at LEP at centre-of-mass energies root(s)=183-209GeV with a total
integrated luminosity of 684.8pb^-1. The cross section of the gamma gamma* ->
rho+ rho- process is determined as a function of the photon virtuality, Q^2,
and the two-photon centre-of-mass energy, W_gg, in the kinematic region:
0.2GeV^2 < Q^2 <0.85GeV^2 and 1.1GeV < W_gg < 3GeV. These results, together
with previous L3 measurements of rho0 rho0 and rho+ rho- production, allow a
study of the gamma gamma* -> rho rho process over the Q^2-region 0.2GeV^2 < Q^2
< 30 GeV^2
Bose-Einstein Correlations of Neutral and Charged Pions in Hadronic Z Decays
Bose-Einstein correlations of both neutral and like-sign charged pion pairs
are measured in a sample of 2 million hadronic Z decays collected with the L3
detector at LEP. The analysis is performed in the four-momentum difference
range 300 MeV < Q < 2 GeV. The radius of the neutral pion source is found to be
smaller than that of charged pions. This result is in qualitative agreement
with the string fragmentation model
Search for Neutral Higgs Bosons of the Minimal Supersymmetric Standard Model in e+e- Interactions at root(s)=192-202GeV
A search for the lightest neutral CP-even and the neutral CP-odd Higgs bosons
of the Minimal Supersymmetric Standard Model is performed using 233.2 pb-1 of
integrated luminosity collected with the L3 detector at LEP at centre-of-mass
energies 192-202 GeV. No signal is observed and lower mass limits are given as
a function of tan(beta) for two scalar top mixing hypotheses. For tan(beta)
greater than 0.8, they are mh > 83.4 GeV and mA > 83.8 GeV at 95 % confidence
level
Measurement of Exclusive rho^0 rho^0 Production in Mid-Virtuality Two-Photon Interactions at LEP
Exclusive rho^0 rho^0 production in two-photon collisions between a
quasi-real and a mid-virtuality photon is studied with data collected at LEP at
centre-of-mass energies 183GeV < sqrt{s} < 209GeV with a total integrated
luminosity of 684.8/pb. The cross section of the process gamma gamma* -> rho^0
rho^0 is determined as a function of the photon virtuality, q^2, and the
two-photon centre-of-mass energy, Wgg, in the kinematic region: 0.2GeV^2 < q^2
< 0.85GeV^2 and 1.1GeV < Wgg < 3GeV
Measurement of Bose-Einstein Correlations in e+e- -> W+W- at root(s)=189GeV
We investigate Bose-Einstein correlations (BEC) in W-pair production at
root(s)=189GeV using the L3 detector at LEP. We observe BEC between particles
from a single W decay in good agreement with those from a light-quark Z decay
sample. We investigate their possible existence between particles coming from
different W's. No evidence for such inter-W BEC is found
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