C9orf72-derived arginine-rich poly-dipeptides impede phase modifiers

Nuclear import receptors (NIRs) not only transport RNA-binding proteins (RBPs) but also modify phase transitions of RBPs by recognizing nuclear localization signals (NLSs). Toxic arginine-rich poly-dipeptides from C9orf72 interact with NIRs and cause nucleocytoplasmic transport deficit. However, the molecular basis for the toxicity of arginine-rich poly-dipeptides toward NIRs function as phase modifiers of RBPs remains unidentified. Here we show that arginine-rich poly-dipeptides impede the ability of NIRs to modify phase transitions of RBPs. Isothermal titration calorimetry and size-exclusion chromatography revealed that proline:arginine (PR) poly-dipeptides tightly bind karyopherin-β2 (Kapβ2) at 1:1 ratio. The nuclear magnetic resonances of Kapβ2 perturbed by PR poly-dipeptides partially overlapped with those perturbed by the designed NLS peptide, suggesting that PR poly-dipeptides target the NLS binding site of Kapβ2. The findings offer mechanistic insights into how phase transitions of RBPs are disabled in C9orf72-related neurodegeneration

Reliability and Validity of Caregivers’ Fear of Falling Index When Caring for Home-Based Rehabilitation Patients With Fall-Related Fractures

Objective To evaluate the reliability and validity of this new measure, called the caregivers’ fear of falling index (CFFI). Methods The study surveyed home-based rehabilitation patients with fall-related fracture, and their primary caregivers. The characteristics of these patients were evaluated, and the caregivers were surveyed using the CFFI and Falls Efficacy Scale-International (FES-I). The reliability of the CFFI was assessed using item-total correlation, while the validity of the CFFI was evaluated through correlation coefficients calculated between the CFFI and the FES-I. Results The participants were 51 patient-caregiver pairs. The internal consistency of the CFFI showed an alpha coefficient of 0.904. No items were excluded in the corrected item-total correlations. The CFFI showed a moderate correlation with FES-I (r=0.432, p=0.002). Conclusion This study found the CFFI to be a reliable and valid tool for measuring the primary caregivers’ fear. The CFFI may be a useful tool for healthcare professionals to identify and supporting these primary caregivers

DJ-1 binds to mitochondrial complex I and maintains its activity

Parkinson's disease (PD) is caused by neuronal cell death, and oxidative stress and mitochondrial dysfunction are thought to be responsible for onset of PD. DJ-1, a causative gene product of a familial form of Parkinson's disease, PARK7, plays roles in transcriptional regulation and anti-oxidative stress. The possible mitochondrial function of DJ-1 has been proposed, but its exact function remains unclear. In this study, we found that DJ-1 directly bound to NDUFA4 and ND1, nuclear and mitochondrial DNA-encoding subunits of mitochondrial complex I, respectively, and was co-localized with complex I and that complex I activity was reduced in DJ-1-knockdown NIH3T3 and HEK293 cells. These findings suggest that DJ-1 is an integral mitochondrial protein and that DJ-1 plays a role in maintenance of mitochondrial complex I activity

Brain imaging of sequential acquisition using a flexible PET scanner and 3-T MRI: quantitative and qualitative assessment

[Objective] A mobile PET scanner termed flexible PET (fxPET) has been designed to fit existing MRI systems. The purpose of this study was to assess brain imaging with fxPET combined with 3-T MRI in comparison with conventional PET (cPET)/CT. [Methods] In this prospective study, 29 subjects with no visible lesions except for mild leukoaraiosis on whole brain imaging underwent 2-deoxy-2-[¹⁸F]fluoro-D-glucose ([¹⁸F]FDG) cPET/CT followed by fxPET and MRI. The registration differences between fxPET and MRI and between cPET and CT were compared by measuring spatial coordinates. Three-dimensional magnetization-prepared rapid acquisition gradient-echo T1-weighted imaging (T1WI) was acquired. We applied two methods of attenuation correction to the fxPET images: MR-based attenuation correction, which yielded fxPET[MRAC]; and CT-based attenuation correction, which yielded fxPET[CTAC]. The three PET datasets were co-registered to the T1WI. Following subcortical segmentation and cortical parcellation, volumes of interest were placed in each PET image to assess physiological accumulation in the brain. SUV[mean] was obtained and compared between the three datasets. We also visually evaluated image distortion and clarity of fxPET[MRAC]. [Results] Mean misregistration of fxPET/MRI was < 3 mm for each margin. Mean registration differences were significantly larger for fxPET/MRI than for cPET/CT except for the superior margin. There were high correlations between the three PET datasets regarding SUV[mean]. On visual evaluation of image quality, the grade of distortion was comparable between fxPET[MRAC] and cPET, and the grade of clarity was acceptable but inferior for fxPET[MRAC] compared with cPET. [Conclusions] fxPET could successfully determine physiological [¹⁸F]FDG uptake; however, improved image clarity is desirable. In this study, fxPET/MRI at 3-T was feasible for brain imaging

Heat-Induced Conformational Transition Mechanism of Heat Shock Factor 1 Investigated by Tryptophan Probe

A transcriptional regulatory system called heat shock response (HSR) has been developed in eukaryotic cells to maintain proteome homeostasis under various stresses. Heat shock factor-1 (Hsf1) plays a central role in HSR, mainly by upregulating molecular chaperones as a transcription factor. Hsf1 forms a complex with chaperones and exists as a monomer in the resting state under normal conditions. However, upon heat shock, Hsf1 is activated by oligomerization. Thus, oligomerization of Hsf1 is considered an important step in HSR. However, the lack of information about Hsf1 monomer structure in the resting state, as well as the structural change via oligomerization at heat response, impeded the understanding of the thermosensing mechanism through oligomerization. In this study, we applied solution biophysical methods, including fluorescence spectroscopy, nuclear magnetic resonance, and circular dichroism spectroscopy, to investigate the heat-induced conformational transition mechanism of Hsf1 leading to oligomerization. Our study showed that Hsf1 forms an inactive closed conformation mediated by intramolecular contact between leucine zippers (LZs), in which the intermolecular contact between the LZs for oligomerization is prevented. As the temperature increases, Hsf1 changes to an open conformation, where the intramolecular LZ interaction is dissolved so that the LZs can form intermolecular contacts to form oligomers in the active form. Furthermore, since the interaction sites with molecular chaperones and nuclear transporters are also expected to be exposed in the open conformation, the conformational change to the open state can lead to understanding the regulation of Hsf1-mediated stress response through interaction with multiple cellular components

Haem-dependent dimerization of PGRMC1/Sigma-2 receptor facilitates cancer proliferation and chemoresistance

Progesterone-receptor membrane component 1 (PGRMC1/Sigma-2 receptor) is a haem-containing protein that interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 to regulate cancer proliferation and chemoresistance; its structural basis remains unknown. Here crystallographic analyses of the PGRMC1 cytosolic domain at 1.95 Å resolution reveal that it forms a stable dimer through stacking interactions of two protruding haem molecules. The haem iron is five-coordinated by Tyr113, and the open surface of the haem mediates dimerization. Carbon monoxide (CO) interferes with PGRMC1 dimerization by binding to the sixth coordination site of the haem. Haem-mediated PGRMC1 dimerization is required for interactions with EGFR and cytochromes P450, cancer proliferation and chemoresistance against anti-cancer drugs; these events are attenuated by either CO or haem deprivation in cancer cells. This study demonstrates protein dimerization via haem-haem stacking, which has not been seen in eukaryotes, and provides insights into its functional significance in cancer