RUTBC1 Functions as a GTPase-activating Protein for Rab32/38 and Regulates Melanogenic Enzyme Trafficking in Melanocytes

Two cell type-specific Rab proteins, Rab32 and Rab38 (Rab32/38), have been proposed as regulating the trafficking of melanogenic enzymes, including tyrosinase and tyrosinase-related protein 1 (Tyrp1), to melanosomes in melanocytes. Like other GTPases, Rab32/38 function as switch molecules that cycle between a GDP-bound inactive form and a GTP-bound active form; the cycle is thought to be regulated by an activating enzyme, guanine nucleotide exchange factor (GEF), and an inactivating enzyme, GTPase-activating protein (GAP), which stimulates the GTPase activity of Rab32/38. Although BLOC-3 has already been identified as a Rab32/38-specific GEF that regulates the trafficking of tyrosinase and Tyrp1, no physiological GAP for Rab32/38 in melanocytes has ever been identified, and it has remained unclear whether Rab32/38 is involved in the trafficking of dopachrome tautomerase, another melanogenic enzyme, in mouse melanocytes. In this study we investigated RUTBC1, which was originally characterized as a Rab9-binding protein and GAP for Rab32 and Rab33B in vitro, and the results demonstrated that RUTBC1 functions as a physiological GAP for Rab32/38 in the trafficking of all three melanogenic enzymes in mouse melanocytes. The results of this study also demonstrated the involvement of Rab9A in the regulation of the RUTBC1 localization and in the trafficking of all three melanogenic enzymes. We discovered that either excess activation or inactivation of Rab32/38 achieved by manipulating RUTBC1 inhibits the trafficking of all three melanogenic enzymes. These results collectively indicate that proper spatiotemporal regulation of Rab32/38 is essential for the trafficking of all three melanogenic enzymes in mouse melanocytes

Proton Irradiation Experiment for the X-ray Charge-Coupled Devices of the Monitor of All-sky X-ray Image mission onboard the International Space Station: I. Experimental Setup and Measurement of the Charge Transfer Inefficiency

We have investigated the radiation damage effects on a CCD to be employed in the Japanese X-ray astronomy mission including the Monitor of All-sky X-ray Image (MAXI) onboard the International Space Station (ISS). Since low energy protons release their energy mainly at the charge transfer channel, resulting a decrease of the charge transfer efficiency, we thus focused on the low energy protons in our experiments. A 171 keV to 3.91 MeV proton beam was irradiated to a given device. We measured the degradation of the charge transfer inefficiency (CTI) as a function of incremental fluence. A 292 keV proton beam degraded the CTI most seriously. Taking into account the proton energy dependence of the CTI, we confirmed that the transfer channel has the lowest radiation tolerance. We have also developed the different device architectures to reduce the radiation damage in orbit. Among them, the ``notch'' CCD, in which the buried channel implant concentration is increased, resulting in a deeper potential well than outside, has three times higher radiation tolerance than that of the normal CCD. We then estimated the charge transfer inefficiency of the CCD in the orbit of ISS, considering the proton energy spectrum. The CTI value is estimated to be 1.1e-5 per each transfer after two years of mission life in the worse case analysis if the highest radiation-tolerant device is employed. This value is well within the acceptable limit and we have confirmed the high radiation-tolerance of CCDs for the MAXI mission.Comment: 17 pages, 2 table, 12 figures. Accepted for publication of Japanese Journal of Applied Physics. High resolution file is available from http://wwwxray.ess.sci.osaka-u.ac.jp/~miyata/paper/proton_cti.pd