The origin and evolution of G protein-coupled receptor kinases

G protein-coupled receptor (GPCR) kinases (GRKs) play key role in homologous desensitization of GPCRs. GRKs phosphorylate activated receptors, promoting high affinity binding of arrestins, which precludes G protein coupling. Direct binding to active GPCRs activates GRKs, so that they selectively phosphorylate only the activated form of the receptor regardless of the accessibility of the substrate peptides within it and their Ser/Thr-containing sequence. Mammalian GRKs were classified into three main lineages, but earlier GRK evolution has not been studied. Here we show that GRKs emerged at the early stages of eukaryotic evolution via an insertion of a kinase similar to ribosomal protein S6 kinase into a loop in RGS domain. GRKs in Metazoa fall into two clades, one including GRK2 and GRK3, and the other consisting of all remaining GRKs, split into GRK1-GRK7 lineage and GRK4-GRK5-GRK6 lineage in vertebrates. One representative of each of the two ancient clades is found as early as placozoan Trichoplax adhaerens. Several protists, two oomycetes and unicellular brown algae have one GRK-like protein, suggesting that the insertion of a kinase domain into the RGS domain preceded the origin of Metazoa. The two GRK families acquired distinct structural units in the N- and C-termini responsible for membrane recruitment and receptor association. Thus, GRKs apparently emerged before animals and rapidly expanded in true Metazoa, most likely due to the need for rapid signalling adjustments in fast-moving animals.     

Reliability and external validity of the six scales of 72-item Sleep-Wake Pattern Assessment Questionnaire (SWPAQ)

The 72-item Sleep-Wake Pattern Assessment Questionnaire (SWPAQ) provides possibility to self-assess individual variation along as many as 6 factorial dimensions. We examined reliability and external validity of each of its 12-item scales, E, M, W, V, F, and S (evening and morning lateness, anytime and daytime wakeability, anytime and night-time sleepability, respectively). Questionnaire data were collected from residents of Novosibirsk and two smaller Russian cities (N = 755 and 720, respectively). Analysis of these two data-sets suggested good and acceptable internal consistency of each of the scales, respectively. Evidence for external validity of each of the 6 scales was obtained by applying stepwise linear regression analysis to data collected from 160 participants of sleep deprivation experiments who were asked to self-report 6 characteristics of their sleep for a week prior to the experiment. As expected, predictors of scales? scores were a later self-reported time for going to bed (E, W), an earlier/later time for final awakening (W/M), a shorter/longer sleep latency (F, S/E), a shorter/longer total sleep time (W + V/F), a lower/higher sleep satisfaction score (M/W + V), a lower/higher nap frequency (V/F), and a lower self-scored sleepiness after deprivation from sleep for a night (W, V).     

Rotational restriction of nascent peptides as an essential element of co-translational protein folding: possible molecular players and structural consequences

Background

A basic tenet of protein science is that all information about the spatial structure of proteins is present in their sequences. Nonetheless, many proteins fail to attain native structure upon experimental denaturation and refolding in vitro, raising the question of the specific role of cellular machinery in protein folding in vivo. Recently, we hypothesized that energy-dependent twisting of the protein backbone is an unappreciated essential factor guiding the protein folding process in vivo. Torque force may be applied by the ribosome co-translationally, and when accompanied by simultaneous restriction of the rotational mobility of the distal part of the growing chain, the resulting tension in the protein backbone would facilitate the formation of local secondary structure and direct the folding process.

Results

Our model of the early stages of protein folding in vivo postulates that the free motion of both terminal regions of the protein during its synthesis and maturation is restricted. The long-known but unexplained phenomenon of statistical overrepresentation of protein termini on the surfaces of the protein structures may be an indication of the backbone twist-based folding mechanism; sustained maintenance of a twist requires that both ends of the protein chain are anchored in space, and if the ends are released only after the majority of folding is complete, they are much more likely to remain on the surface of the molecule. We identified the molecular components that are likely to play a role in the twisting of the nascent protein chain and in the anchoring of its N-terminus. The twist may be induced at the C-terminus of the nascent polypeptide by the peptidyltransferase center of the ribosome. Several ribosome-associated proteins, including the trigger factor in bacteria and the nascent polypeptide-associated complex in archaea and eukaryotes, may restrict the rotational mobility of the N-proximal regions of the peptides.

Conclusions

Many experimental observations are consistent with the hypothesis of co-translational twisting of the protein backbone. Several molecular players in this hypothetical mechanism of protein folding can be suggested. In addition, the new view of protein folding in vivo opens the possibility of novel potential drug targets to combat human protein folding diseases.

Reviewers

This article was reviewed by Lakshminarayan Iyer and István Simon.

     

Antidepressant effects of light therapy and "natural" treatments for winter depression

Although bright light treatment may alleviate the symptoms of winter depression, it still remains to be clarified whether chronobiological mechanisms are involved in this antidepressant response. We studied the therapeutic action of bright light in 61 women with and 36 women without winter depression at the medical academic hospital near Novosibirsk (55 degrees North). Bright light was administered with cool-white incandescent lamp for seven days, two hours daily. The treatment started from either 8:00 (n = 29 patients and 16 controls) or 16:00 (n = 24 and 14, respectively) or 18:00 (n = 8 and 6, respectively). The subsets of bright light-treated subjects were then restudied in wintertime before and after one-week vacation in Firuza resort (south of Turkmeniya, 38 degrees North) (n = 19 and 0, respectively), in summertime (n = 42 and 18, respectively) and in the next winter before and after a week 30-min exposure in the morning hours to dim red light emitting “Light Cap” (n = 9 and 0, respectively). The results suggest that, in controls, mood slightly but statistically significantly improved after light treatment and in summer. In patients, the improvement of mood after one week of bright light was comparable with the effects of such “natural” treatments as trips south and transition from winter to summer seasons. Although next winter response to 0.5-h dim light was clinically significant, it was significantly worse compared to the previous response to 2-h bright light. Our therapeutic results indicate that, despite the different potential phase-shifting effect of bright light administered in the morning and in the second half of the day, the responses to all treatments are equally beneficial. This finding provides evidence against the view that circadian phase shifts are the key to the pathogenesis of winter depression and efficacy of light therapy. Although several different physiological effects of light therapy might be involved in the antidepressant response, none of them seems to be of more importance compared to psychological components of this response. Ours and earlier published reports on the independence of beneficial action of bright light from treatment timing support the suggestion that, in the open investigational trials, the placebo effect accounts for a large portion of the antidepressant response. We also reviewed several facts pointing to the close dependence of antidepressant effects of non-drug therapy upon patients' expectations and researchers' enthusiasm. In sum, unlike patients' chronobiology, their psychology seems to be most powerful mediator of the clinical response to bright light.     

Evolutionary perspective on innate immune recognition

Analysis of human and Drosophila genomes demonstrates an ancient origin of innate immunity and the diversity of the mechanisms of innate immune recognition.     

Menstrual Phase Response to Nocturnal Light

The aims of the study were to test whether nocturnal white light can normalize menstrual cycles in oligomenorrheic women, and whether the phase of the menstrual cycle in which light is given is important for the shortening effect. Twenty-five women with long menstrual cycles (35.9–53.4 days on average) were treated for 1–3 cycles, each of which was preceded and followed by at least two untreated cycles. Treatments were 100 watt bedside lights administered for 5 consecutive nights. They centered at three different phases of the menstrual cycle: 6–7th, 14–17th or 23–25th days of the treated cycle (early, middle or late treatment, respectively). On average, the treatment cycle lengths were modestly, but significantly reduced compared to the duration of baseline cycles (more than 11 %). The difference in the effects of the early, middle and late treatment was not significant. However, if middle or late treatments were administered in the latter half of the interval between the menstrual cycle onset and probable time of ovulation, reductions of the treated cycle length were substantial (more than 20 %, resulting in cycles less than 33 days on average; p < 0.001). Other treatments produced only weak (up to 7 %), if any, cycle reductions. Moreover, we found a strong correlation (p < 0.001) between the duration of baseline cycle and differential effect of middle treatment (compared to early or late treatment). Middle treatments reduced treated cycle duration to the normal range in the subjects with shorter mean baseline cycles (<42 days), while in the subjects with longer duration of baseline cycle the shortening effect was produced by late treatments (p = 0.005 and p = 0.001, respectively). The results support the suggestion that a bedside lamp used on nights prior to ovulation can cause reduction of long menstrual cycles.     

Unevenness of distribution of historical events throughout an 11-year solar cycle]

Tchizhevsky hypothesis (1922) of historical process heliotaraxia (helios--sun, taracsio--perturb) was empirically tested. Samples of near 13 and 4.6 thousand events mentioned in Chronology sections of two largest Soviet historical handbooks were analyzed. Events were classified into 4 groups on the basis of "strength" and "social contradictions meaning" of their names, called tolerance and polarity: tolerant--intolerant (e.f. riot--roform) and polar--neutral (e.f. civil war-external war). It was found that frequency and polarity of historical events increased in maximum of sunspot cycle and in the next year as compared with minimum and the year before minimum. The probability of revolution (the most polar and intolerant name of historical event) is the highest in maximum and the lowest in the year before minimum. Intolerance of polar events increased and neutral events decreased in maximum. All these relations were highly significant (P < 0.001). It was concluded that heliotaraxic phenomena exist and are basically associated with year of sunspot maximum.     

Identification of Elongin C and Skp1 sequences that determine Cullin selection

The multiprotein von Hippel-Lindau (VHL) tumor suppressor and Skp1-Cul1-F-box protein (SCF) complexes belong to families of structurally related E3 ubiquitin ligases. In the VHL ubiquitin ligase, the VHL protein serves as the substrate recognition subunit, which is linked by the adaptor protein Elongin C to a heterodimeric Cul2/Rbx1 module that activates ubiquitylation of target proteins by the E2 ubiquitin-conjugating enzyme Ubc5. In SCF ubiquitin ligases, F-box proteins serve as substrate recognition subunits, which are linked by the Elongin C-like adaptor protein Skp1 to a Cul1/Rbx1 module that activates ubiquitylation of target proteins, in most cases by the E2 Cdc34. In this report, we investigate the functions of the Elongin C and Skp1 proteins in reconstitution of VHL and SCF ubiquitin ligases. We identify Elongin C and Skp1 structural elements responsible for selective interaction with their cognate Cullin/Rbx1 modules. In addition, using altered specificity Elongin C and F-box protein mutants, we investigate models for the mechanism underlying E2 selection by VHL and SCF ubiquitin ligases. Our findings provide evidence that E2 selection by VHL and SCF ubiquitin ligases is determined not solely by the Cullin/Rbx1 module, the target protein, or the integrity of the substrate recognition subunit but by yet to be elucidated features of these macromolecular complexes.     

Phase Typing of Patients with Seasonal Affective Disorder: A Test for the Phase Shift Hypothesis

The study evaluates the phase-shift hypothesis for seasonal affective disorder (Lewy et al., 1987, 1988) in parallel-design comparison of effects of morning (800-1000) or afternoon (1600-1800) light treatment on mood and circadian phase. Subjective arousal, body temperature, melatonin and cortisol were measured at 800, 1200, 1600, 2000 and 2400 in 23 women with seasonal depression and 20 controls before and after a week of bright light (2 hours per day). The rates of clinical response to both treatments were similar. Comparison of circadian variations did not provide evidence for significant phase-delay in patients compared to controls. However, morning light produced significant phase advance in patients, but not in controls. Also we found that advance phase shifts in well-responded patients were more often than in patients with worse response and controls. Before light treatment phase concordance between different variables in patients was lower compared to either themselves after light treatment or controls before and after light treatment. Dependence of antidepressant response to light from pretreatment circadian phases was also observed. Those patients who responded worse to morning light tended to have advance circadian phases, while those who responded worse to afternoon light tended to have delay phases. Although some results are lending support for the phase-shift hypothesis, other explanations for mechanisms by which biological rhythms are implicated in winter depression and light treatment might be suggested.