Cryo-EM structures of the D290V mutant of the hnRNPA2 low-complexity domain suggests how D290V affects phase separation and aggregation.

Heterogeneous nuclear ribonucleoprotein A2 (hnRNPA2) is a human ribonucleoprotein that transports RNA to designated locations for translation via its ability to phase separate. Its mutated form, D290V, is implicated in multisystem proteinopathy known to afflict two families, mainly with myopathy and Paget's disease of bone. Here, we investigate this mutant form of hnRNPA2 by determining cryo-EM structures of the recombinant D290V low complexity domain. We find that the mutant form of hnRNPA2 differs from the WT fibrils in four ways. In contrast to the WT fibrils, the PY-nuclear localization signals in the fibril cores of all three mutant polymorphs are less accessible to chaperones. Also, the mutant fibrils are more stable than WT fibrils as judged by phase separation, thermal stability, and energetic calculations. Similar to other pathogenic amyloids, the mutant fibrils are polymorphic. Thus, these structures offer evidence to explain how a D-to-V missense mutation diverts the assembly of reversible, functional amyloid-like fibrils into the assembly of pathogenic amyloid, and may shed light on analogous conversions occurring in other ribonucleoproteins that lead to neurological diseases such as amyotrophic lateral sclerosis and frontotemporal dementia.

Prevalence and species distribution of the low-complexity, amyloid-like, reversible, kinked segment structural motif in amyloid-like fibrils.

Membraneless organelles (MLOs) are vital and dynamic reaction centers in cells that compartmentalize the cytoplasm in the absence of a membrane. Multivalent interactions between protein low-complexity domains contribute to MLO organization. Previously, we used computational methods to identify structural motifs termed low-complexity amyloid-like reversible kinked segments (LARKS) that promote phase transition to form hydrogels and that are common in human proteins that participate in MLOs. Here, we searched for LARKS in the proteomes of six model organisms: Homo sapiens, Drosophila melanogaster, Plasmodium falciparum, Saccharomyces cerevisiae, Mycobacterium tuberculosis, and Escherichia coli to gain an understanding of the distribution of LARKS in the proteomes of various species. We found that LARKS are abundant in M. tuberculosis, D. melanogaster, and H. sapiens but not in S. cerevisiae or P. falciparum. LARKS have high glycine content, which enables kinks to form as exemplified by the known LARKS-rich amyloidogenic structures of TDP43, FUS, and hnRNPA2, three proteins that are known to participate in MLOs. These results support the idea of LARKS as an evolved structural motif. Based on these results, we also established the LARKSdb Web server, which permits users to search for LARKS in their protein sequences of interest.

DNAJB8 oligomerization is mediated by an aromatic-rich motif that is dispensable for substrate activity.

J-domain protein (JDP) molecular chaperones have emerged as central players that maintain a healthy proteome. The diverse members of the JDP family function as monomers/dimers and a small subset assemble into micron-sized oligomers. The oligomeric JDP members have eluded structural characterization due to their low-complexity, intrinsically disordered middle domains. This in turn, obscures the biological significance of these larger oligomers in protein folding processes. Here, we identified a short, aromatic motif within DNAJB8 that drives self-assembly through π-π stacking and determined its X-ray structure. We show that mutations in the motif disrupt DNAJB8 oligomerization in vitro and in cells. DNAJB8 variants that are unable to assemble bind to misfolded tau seeds more specifically and retain capacity to reduce protein aggregation in vitro and in cells. We propose a new model for DNAJB8 function in which the sequences in the low-complexity domains play distinct roles in assembly and substrate activity.

Chronotype, Social Jetlag, and Nicotine Use.

Late chronotype, which often leads to higher social jetlag (SJL), is strongly associated with the prevalence of smoking. Any circadian disruption, strain, or misalignment, results in people not being able to live according to their biological time as is described by SJL, which we will therefore use as umbrella term. We hypothesized two scenarios potentially explaining the association between smoking and SJL: (A) If smoking delays the clock, circadian phase should advance upon quitting. (B) If people smoke more to compensate the consequences of SJL, circadian phase should not change upon quitting. To distinguish between these two hypotheses, we accompanied participants of a smoking cessation program (not involving nicotine replacement products) across the cessation intervention (3 weeks prior and 6 weeks after) by monitoring their circadian behavior, sleep quality, and daytime sleepiness via questionnaires and actimetry. Our results show no effects of cessation on SJL, chronotype, sleep quality, or daytime sleepiness, thereby favoring scenario (B). Thus, smoking may be a consequence of rather than a cause for SJL. Daytime sleepiness was a significant predictor for the outcome in our model but did not improve with cessation.

The Relationship Among Chronotype, Hardiness, Affect, and Talent and Their Effects on Performance in a Military Context.

Individual preference for morning or evening activities (chronotype), affect, hardiness, and talent are associated with a variety of performance outcomes. This longitudinal study was designed to investigate the degree to which these variables are associated with academic, physical, and military performance. Self-reported measures of chronotype, affect, and hardiness were collected from 1149 cadets from the Class of 2016 upon entry to the United States Military Academy. Talent, a composite of academic, leadership, and physical fitness scores were drawn from cadet records. Academic, military, and physical performance measures were collected at graduation 4 years later. The results indicated that a morning orientation was associated with better physical and military performance. Higher talent scores, as well as lower levels of negative affect, were associated with better performance across all three performance measures. Hardiness was only associated with military performance. The findings suggest that a morning orientation and less negative affect may result in better performance overall within a challenging and structured military environment. Future studies of chronotype shifts may provide further insight into associated performance benefits.

Aspects of the breeding ecology and behaviors of the Bar-tailed Lark (Ammomanes cinctura) from Ha'il region in north of Saudi Arabia.

The Bar-tailed Lark (Ammomanes cinctura) breeds in desert and semi-desert areas of the Saharo-Sindian region, from north-west Africa through the arid plains of the Arabian Peninsula to the Sind. Despite having a wide distribution, little information is available on the breeding ecology of this species. This study was conducted in a desert in the north of Saudi Arabia, where the daytime ambient temperature may exceed 40 °C. In contrast, the night ambient temperature may reach less than 10 °C in late spring and early summer. The objectives of this study were to collect some baseline data on some aspects of the breeding ecology of this species and to record the nest attendance behavior. The study found that Bar-tailed Larks preferred to nest under shrub trunks, which may camouflage both nests and incubating parents against predators and protect eggs, nestlings and incubating parents from hostile weather conditions. Moreover, nest attendance was high, as Bar-tailed Lark parents incubated their eggs 95.97 ± 2.62% over the entire day, and they seemed to maintain the eggs at temperatures around 23-33 °C. In addition, they incubated more at night than during the daytime. Temperatures under the shrubs at night fell below 21 °C, thus parents increased the nest attendance to warm the eggs and prevent the embryos from exposure to lethal temperatures.

Karyopherins and condensates.

Several aggregation-prone RNA-binding proteins, including FUS, EWS, TAF15, hnRNP A1, hnRNP A2, and TDP-43, are mutated in neurodegenerative diseases. The nuclear-cytoplasmic distribution of these proteins is controlled by proteins in the karyopherin family of nuclear transport factors (Kaps). Recent studies have shown that Kaps not only transport these proteins but also inhibit their self-association/aggregation, acting as molecular chaperones. This chaperone activity is impaired for disease-causing mutants of the RNA-binding proteins. Here, we review physical data on the mechanisms of self-association of several disease-associated RNA-binding proteins, through liquid-liquid phase separation and amyloid fiber formation. In each case, we relate these data to biophysical, biochemical, and cell biological data on the inhibition of self-association by Kaps. Our analyses suggest that Kaps may be effective chaperones because they contain large surfaces with diverse physical properties that enable them to engage multiple different regions of their cargo proteins, blocking self-association.