166 research outputs found
Functional Analysis of the SPA Gene Family in Arabidopsis thaliana
Ambient light conditions affect development throughout the plant life cycle, including seed germination, seedling development and the induction of flowering. In the model plant Arabidopsis, the COP1-SPA ubiquitin ligase complex plays a central role in suppressing light signaling in darkness. The COP1-SPA complex targets positively acting factors like HY5, a protein necessary for normal seedling development in the light, several photoreceptors and the flowering time regulator CONSTANS for degradation via the 26S proteasome. Therefore, one of the major functions of the light signal transduction pathways is the inactivation of the COP1-SPA complex. While COP1 is a single copy gene, the SPA proteins are encoded by four different loci (SPA1-SPA4). All SPA proteins have redundant, but also distinct functions in regulating plant development. SPA1 and SPA2 are the key regulators that suppress photomorphogenesis in dark-grown seedlings. Over-stimulation in light-grown seedlings is primarily prevented by SPA1, and to a minor extent, also by SPA3 and SPA4. SPA2, in contrast has only negligible function in the light. SPA1 is sufficient for repressing flowering under non-inductive short-day conditions. Here, I show that distinct functions of the SPA genes partially correlate with their distinct gene expression patterns. RNA gel blot-analysis revealed that the expression of SPA1, SPA3 and SPA4, but not that of SPA2, is positively influenced by light of different wavelengths. All main photoreceptors contribute to the up-regulation of these SPA transcripts, implying that photoreceptors initiate a negative feedback regulation, which might protect plants from over-stimulation by light. GUS reporter gene experiments show that SPA genes exhibit somewhat distinct tissue-specific expression patterns, which might be important for tissue specific regulation of COP1-SPA targets. However, differences in SPA gene expression cannot account for all distinct SPA gene functions. Promoter-swap experiments with SPA1, SPA2 and SPA4 show that all SPA proteins are potent repressors in dark-grown seedlings. SPA1 and SPA4 also act as repressor in the light. SPA2, however, can never act as a repressor in the light, not even when it is expressed from the strong light-induced SPA1 promoter. These results show that SPA proteins themselves feature properties that determine characteristic SPA protein functions. All SPA proteins feature a characteristic domain structure with a C-terminal WD-repeat, a central coiled-coil domain and a less well-conserved N-terminus that includes a kinase-like motif. The WD-repeat domain and the coiled-coil domain are essential for formation of the COP1-SPA complex as well as interactions with various ubiquitination targets. In contrast, the function of the N-terminal domain is unknown. Aiming to determine the importance of the N-terminal domain of SPA1, I conducted a structure-function analysis. While the N-terminal domain of SPA1 is dispensable for SPA1 function in the seedling stage, this domain is required for SPA1-mediated repression of flowering in non-inductive short-day conditions. These results indicate, that the SPA1 N-terminal domain can full-fill an essential function
Recommended from our members
Optical field ionization of atoms and ions using ultrashort laser pulses
This dissertation research is an investigation of the strong optical field ionization of atoms and ions by 120-fs, 614-run laser pulses and 130-fs, 800-nm laser pulses. The experiments have shown ionization that is enhanced above the predictions of sequential tunneling models for He{sup +2}, Ne{sup +2} and Ar{sup +2}. The ion yields for He{sup +l}, Ne{sup +l} and Ar{sup +l} agree well with the theoretical predictions of optical tunneling models. Investigation of the polarization dependence of the ionization indicates that the enhancements are consistent with a nonsequential ionization mechanism in which the linearly polarized field drives the electron wavefunction back toward the ion core and causes double ionization through inelastic e-2e scattering. These investigations have initiated a number of other studies by other groups and are of current scientific interest in the fields of high-irradiance laser-matter interactions and production of high-density plasmas. This work involved: (1) Understanding the characteristic nature of the ion yields produced by tunneling ionization through investigation of analytic solutions for tunneling at optical frequencies. (2) Extensive characterization of the pulses produced by 614-nm and 800-ran ultrashort pulse lasers. Absolute calibration of the irradiance scale produced shows the practicality of the inverse problem--measuring peak laser irradiance using ion yields. (3) Measuring the ion yields for three noble gases using linear, circular and elliptical polarizations of laser pulses at 614-nm and 800-nm. The measurements are some of the first measurements for pulse widths as low as 120-fs
High-efficiency temporally decorrelated multifocal arrays for multiphoton microscopy and micromachining
Temporally decorrelated muItifocal array for high-speed, high-resolution, muItiphoton imaging and micromachining
Recommended from our members
Spatial and Temporal Characterization of Femtosecond Pulses at High-Numerical Aperture Using Collinear, Background-Free, Third-Harmonic Autocorrelation
We show that a simple plane wave analysis can be used even under tight focusing conditions to predict the dependence of third-harmonic generation on the polarization state of the incident beam. Exploiting this fact, we then show that circularly polarized beams may be used to spatially characterize the beam focus and temporally characterize ultrashort pulses in high numerical aperture systems by experimentally demonstrating, for the first time, novel collinear, background-free, third-harmonic intensity autocorrelations in time and space in a high numerical aperture microscope. We also discuss the possibility of using third harmonic generation with circularly polarized beams for background-free collinear frequency resolved optical gating
Recommended from our members
Double ionization in gases driven by optical field ionization using ultrashort laser pulses
Rapid optical field ionization (OFI) for atoms and ions is distinguished using ultra-short laser pulses focused to peak fields comparable to Coulomb binding values. This nonperturbative field description facilitates understanding of standard sequential ionization and distinguishes the double (or two-electron ejection) ionization process. Following our discovery of double optical field ionization in He{sup +2} at 614 nm we have extended experiments to include helium, neon and argon at an 800 nm wavelength with varying ellipticity. We compare double ionization data with predictions from the semi-classical model by Corkum. New experimental results at 800 nm use 130 femtosecond laser pulses focused to intensities of order 10{sup 16} W/cm in the extraction region of a 1 meter time-of-flight spectrometer. Gas pressures ranged from 10{sup {minus}8} to 10{sup {minus}5} torr
Recommended from our members
Hyperdispersion Grating Arrangements for Compact Pulse Compressors and Expanders
A novel, but general, arrangement of parallel sets of gratings is presented that can effectively increase the dispersion of pulse compressors and expanders by over an order of magnitude. These arrangements will dramatically reduce the footprint of the pulse compressors and expanders used in CPA
80 Gb/s optimised pulse source using a gain-switched laser diode in conjunction with a nonlinearly chirped grating
The authors demonstrate the generation of transforms limited short optical pulses, which display excellent spectral and temporal qualities by employing a novel technology, based on an externally injected gain-switched laser in conjunction with a non-linearly chirped grating. Using this technique, 3.5 ps optical pulses, exhibiting a time bandwidth product of 0.45, are generated, which are suitable for use in high-speed 80 Gb/s OTDM communications systems
Consistency and variability in the behavior of mature, isolation-reared, male rhesus macaques
- …
