RESUMO
Stress granules (SGs) are sites for mRNA storage, protection, and translation repression. TIA1 and TIAR1 are two RNA-binding proteins that are key players in SGs formation in mammals. TIA1/TIAR have a prion-like domain (PrD) in their C-terminal that promotes liquid-phase separation. Lack of any TIA1/TIAR has severe consequences in mice. However, it is not clear whether the failure to form proper SGs is the cause of any of these problems. We disrupted two predicted α-helices within the prion-like domain of the Caenohabditis elegans TIA1/TIAR homolog, TIAR-1, to test whether its association with SGs is important for the nematode. We found that tiar-1 PrD mutant animals continued to form TIAR-1 condensates under stress in the C. elegans gonad. Nonetheless, TIAR-1 condensates appeared fragile and disassembled quickly after stress. Apparently, the SGs continued to associate regularly as observed with CGH-1, an SG marker. Like tiar-1-knockout nematodes, tiar-1 PrD mutant animals exhibited fertility problems and a shorter lifespan. Notwithstanding this, tiar-1 PrD mutant nematodes were no sensitive to stress. Our data demonstrate that the predicted prion-like domain of TIAR-1 is important for its association with stress granules. Moreover, this domain may also play a significant role in various TIAR-1 functions unrelated to stress, such as fertility, embryogenesis and lifespan.
RESUMO
Photoreflectance-difference (PR/PRD) and reflectance-difference (RD) spectroscopies employ synchronic detection usually with lock-in amplifiers operating at moderate (200-1000 Hz) and high (50-100 KHz) modulation frequencies, respectively. Here, we report a measurement system for these spectroscopies based on a multichannel CCD spectrometer without a lock-in amplifier. In the proposed scheme, a typical PRD or RD spectrum consists of numerical subtractions between a thousand CCD captures recorded, while a photoelastic modulator is either operating or inhibited. This is advantageous and fits the slow response of CCD detectors to high modulation frequencies. The resulting spectra are processed with Savitzky-Golay filtering and compared well with those measured with conventional scanning systems based on lock-in amplifiers.
RESUMO
A kinetic description of Alfvén-cyclotron magnetic fluctuations for anisotropic electron-proton quasistable plasmas is studied. An analytical treatment, based on the fluctuation-dissipation theorem, consistently shows that spontaneous fluctuations in plasmas with stable distributions significantly contribute to the observed magnetic fluctuations in the solar wind, as seen, for example, in [S. D. Bale et al., Phys. Rev. Lett. 103, 211101 (2009)], even far below from the instability thresholds. Furthermore, these results, which do not require any adjustable parameters or wave excitations, are consistent with the results provided by hybrid simulations. It is expected that this analysis contributes to our understanding of the nature of magnetic fluctuations in the solar wind.
RESUMO
Apoptosis is an important mechanism for maintaining germ line health. In Caenorhabditis elegans, germ cell apoptosis occurs under normal conditions to sustain gonad homeostasis and oocyte quality. Under stress, germ cell apoptosis can be triggered via different pathways, including the following: (i) the CEP-1/p53 pathway, which induces germ cell apoptosis when animals are exposed to DNA damage; (ii) the mitogen-activated protein kinase kinase (MAPKK) pathway, which triggers germ cell apoptosis when animals are exposed to heat shock, oxidative stress, or osmotic stress; and (iii) an unknown mechanism that triggers germ cell apoptosis during starvation. Here, we address how starvation induces germ cell apoptosis. Using polysomal profiling, we found that starvation for 6 h reduces the translationally active ribosomes, which differentially affect the mRNAs of the core apoptotic machinery and some of its regulators. During starvation, lin-35/Rb mRNA increases its expression, resulting in the accumulation of this protein. As a consequence, LIN-35 downregulates the expression of the antiapoptotic gene ced-9/Bcl-2. We observed that the reduced translation of ced-9/Bcl-2 mRNA during food deprivation together with its downregulation drastically affects its protein accumulation. We propose that CED-9/Bcl-2 downregulation via LIN-35/Rb triggers germ cell apoptosis in C. elegans in response to starvation.
Assuntos
Apoptose/genética , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/metabolismo , Células Germinativas/patologia , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Repressoras/metabolismo , Estresse Fisiológico , Animais , Proteínas Reguladoras de Apoptose/genética , Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/biossíntese , Proteínas de Ligação ao Cálcio/genética , Caspases/genética , Regulação para Baixo , Fatores de Transcrição E2F/genética , Regulação da Expressão Gênica , Proteínas de Membrana/genética , Biossíntese de Proteínas/genética , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Interferência de RNA , RNA Mensageiro/biossíntese , RNA Interferente Pequeno , Proteínas Repressoras/genética , Ribossomos/metabolismo , Fatores de Transcrição/genéticaRESUMO
We report on a rapid, 32-channel reflectance-difference (RD) spectrometer with sub-second spectra acquisition times and ΔR/R sensitivity in the upper 10(-4) range. The spectrometer is based on a 50 kHz photo-elastic modulator for light polarization modulation and on a lock-in amplifier for signal harmonic analysis. Multichannel operation is allowed by multiplexing the 32 outputs of the spectrometer into the input of the lock-in amplifier. The spectrometer spans a wavelength range of 230 nm that can be tuned to cover E(1) and E(1) + Δ(1) transitions for a number of III-V semiconductors at epitaxial growth temperatures, including GaAs, InAs, AlAs, and their alloys. We present two examples of real-time measurements to demonstrate the performance of the RD spectrometer, namely, the evolution of the RD spectrum of GaAs (001) annealed at 500 °C and the time-dependent RD spectrum during the first stages of the epitaxial growth of In(0.3)Ga(0.7)As on GaAs (001) substrates.
RESUMO
In Caenorhabditis elegans, several distinct apoptosis pathways have been characterized in the germline. The physiological pathway is though to eliminate excess germ cells during oogenesis to maintain gonad homeostasis and it is activated by unknown mechanisms. The DNA damage-induced germ cell apoptosis occurs in response to genotoxic agents and involves the proteins EGL-1 and CED-13, and the DNA damage response protein p53. Germ cell apoptosis can also be induced in response to pathogen infection through an EGL-1 dependent pathway. To gain insight into the mechanism and functions of germ cell apoptosis, we investigated whether and how other forms of stress induce this cell death. We found that oxidative, osmotic, heat shock and starvation stresses induce germ cell apoptosis through a p53 and EGL-1 independent pathway. We also learned that the MAPK kinases MEK-1 and SEK-1, and the p53 antagonist protein ABL-1, are essential for stress-induced germ cell apoptosis. We conclude that in C. elegans responses to various stresses that do not involve genotoxicity include an increase in germ cell apoptosis through the physiological pathway.