The ability to associate with activation domains in vitro is not required for the TATA box-binding protein to support activated transcription in vivo.

The TATA box-binding protein (TBP) interacts in vitro with the activation domains of many viral and cellular transcription factors and has been proposed to be a direct target for transcriptional activators. We have examined the functional relevance of activator-TBP association in vitro to transcriptional activation in vivo. We show that alanine substitution mutations in a single loop of TBP can disrupt its association in vitro with the activation domains of the herpes simplex virus activator VP16 and of the human tumor suppressor protein p53; these mutations do not, however, disrupt the transcriptional response of TBP to either activation domain in vivo. Moreover, we show that a region of VP16 distinct from its activation domain can also tightly associate with TBP in vitro, but fails to activate transcription in vivo. These data suggest that the ability of TBP to interact with activation domains in vitro is not directly relevant to its ability to support activated transcription in vivo.

Point mutation of the autophosphorylation site or in the nuclear location signal causes protein kinase A RII beta regulatory subunit to lose its ability to revert transformed fibroblasts.

The RII beta regulatory subunit of cAMP-dependent protein kinase (PKA) contains an autophosphorylation site and a nuclear location signal, KKRK. We approached the structure-function analysis of RII beta by using site-directed mutagenesis. Ser114 (the autophosphorylation site) of human RII beta was replaced with Ala (RII beta-P) or Arg264 of KKRK was replaced with Met (RII beta-K). ras-transformed NIH 3T3 (DT) cells were transfected with expression vectors for RII beta, RII beta-P, and RII beta-K, and the effects on PKA isozyme distribution and transformation properties were analyzed. DT cells contained PKA-I and PKA-II isozymes in a 1:2 ratio. Over-expression of wild-type or mutant RII beta resulted in an increase in PKA-II and the elimination of PKA-I. Only wild-type RII beta cells demonstrated inhibition of both anchorage-dependent and -independent growth and phenotypic change. The growth inhibitory effect of RII beta overexpression was not due to suppression of ras expression but was correlated with nuclear accumulation of RII beta. DT cells demonstrated growth inhibition and phenotypic change upon treatment with 8-Cl-cAMP. RII beta-P or RII beta-K cells failed to respond to 8-Cl-cAMP. These data suggest that autophosphorylation and nuclear location signal sequences are integral parts of the growth regulatory mechanism of RII beta.

The intrinsic ability of ribosomes to bind to endoplasmic reticulum membranes is regulated by signal recognition particle and nascent-polypeptide-associated complex.

Signal peptides direct the cotranslational targeting of nascent polypeptides to the endoplasmic reticulum (ER). It is currently believed that the signal recognition particle (SRP) mediates this targeting by first binding to signal peptides and then by directing the ribosome/nascent chain/SRP complex to the SRP receptor at the ER. We show that ribosomes can mediate targeting by directly binding to translocation sites. When purified away from cytosolic factors, including SRP and nascent-polypeptide-associated complex (NAC), in vitro assembled translation intermediates representing ribosome/nascent-chain complexes efficiently bound to microsomal membranes, and their nascent polypeptides could subsequently be efficiently translocated. Because removal of cytosolic factors from the ribosome/nascent-chain complexes also resulted in mistargeting of signalless nascent polypeptides, we previously investigated whether readdition of cytosolic factors, such as NAC and SRP, could restore fidelity to targeting. Without SRP, NAC prevented all nascent-chain-containing ribosomes from binding to the ER membrane. Furthermore, SRP prevented NAC from blocking ribosome-membrane association only when the nascent polypeptide contained a signal. Thus, NAC is a global ribosome-binding prevention factor regulated in activity by signal-peptide-directed SRP binding. A model presents ribosomes as the targeting vectors for delivering nascent polypeptides to translocation sites. In conjunction with signal peptides, SRP and NAC contribute to this specificity of ribosomal function by regulating exposure of a ribosomal membrane attachment site that binds to receptors in the ER membrane.

Modification of rhodamine staining allows identification of hematopoietic stem cells with preferential short-term or long-term bone marrow-repopulating ability.

We have developed a modified rhodamine (Rho) staining procedure to study uptake and efflux in murine hematopoietic stem cells. Distinct populations of Rho++ (bright), Rho+ (dull), and Rho- (negative) cells could be discriminated. Sorted Rho- cells were subjected to a second Rho staining procedure with the P-glycoprotein blocking agent verapamil (VP). Most cells became Rho positive [Rho-/Rho(VP)+ cells] and some remained Rho negative [Rho-/Rho(VP)- cells]. These cell fractions were characterized by their marrow-repopulating ability in a syngeneic, sex-mismatch transplantation model. Short-term repopulating ability was determined by recipient survival for at least 6 weeks after lethal irradiation and transplantation--i.e., radioprotection. Long-term repopulating ability at 6 months after transplantation was measured by fluorescence in situ hybridization with a Y-chromosome-specific probe, by graft function and recipient survival. Marrow-repopulating cells were mainly present in the small Rho- cell fraction. Transplantation of 30 Rho- cells resulted in 50% radioprotection and > 80% donor repopulation in marrow, spleen, and thymus 6 months after transplantation. Cotransplantation of cells from both fractions in individual mice directly showed that within this Rho- cell fraction, the Rho-/Rho(VP)+ cells exhibited mainly short-term and the Rho-/Rho(VP)- cells exhibited mainly long-term repopulating ability. Our results indicate that hematopoietic stem cells have relatively high P-glycoprotein expression and that the cells responsible for long-term repopulating ability can be separated from cells exhibiting short-term repopulating ability, probably by a reduced mitochondrial Rho-binding capacity.

Conformation, energy, and folding ability of selected amino acid sequences.

Evolutionary selection of sequences is studied with a knowledge-based Hamiltonian to find the design principle for folding to a model protein structure. With sequences selected by naive energy minimization, the model structure tends to be unstable and the folding ability is low. Sequences with high folding ability have only the low-lying energy minimum but also an energy landscape which is similar to that found for the native sequence over a wide region of the conformation space. Though there is a large fluctuation in foldable sequences, the hydrophobicity pattern and the glycine locations are preserved among them. Implications of the design principle for the molecular mechanism of folding are discussed.

The Impact of Prefrontal Cortex "Warm Up" on Immediate Cognitive Reappraisal Ability in Older Adolescents with Elevated Symptoms of Depression

Cognitive Reappraisal (CR) is a central component of Cognitive Behavioral Therapy for adolescent depression. Yet, previous research indicates that a brain region highly associated with successful CR in adults, the Prefrontal Cortex (PFC), is not fully developed until early adulthood. Thus, there is growing concern that CBT interventions directed at building CR abilities in depressed teens might be constrained by PFC immaturity. However, CR is an effective strategy for regulating affect. The current study evaluated an intervention aimed at enhancing CR performance through PFC “warm up” with a working memory task. Additionally, the study examined moderators of intervention response, as well as cognitive correlates of self-reported CR use. Participants included 48 older adolescents (mean age=19.1, 89% female) with elevated symptoms of depression who were randomly assigned to a lab-based WM or control activity followed by a CR task. Overall, results failed to support the effectiveness of “warm up” to augment CR performance. However, current level of depression predicted negative bias and sadness ratings after CR instructions, and this effect was qualified by an interaction with condition. The moderator analysis showed that depressive symptoms interacted with condition such that in the control condition, participants with higher depressive symptoms had significantly lower negative bias scores than individuals with lower depressive symptoms, but this pattern was not found in the experimental condition. Contrary to hypotheses, history of depression did not moderate treatment response. Additional analyses explored alternative explanations for the lack of intervention effects. There was some evidence to suggest that the WM task was frustrating and cognitively taxing. However, irritation scores and overall WM task accuracy did not predict subsequent CR performance. Lastly, multiple cognitive variables emerged as correlates of self-reported CR use, with cognitive flexibility contributing unique variance to self-reported CR use. Results pointed to new directions for improving CR performance among youth with elevated symptoms of depression.

General mental ability as moderator of personality traits as predictors of early career success

In this paper, we examine the effects of general mental ability (GMA) and the personality traits defined in the big five model on extrinsic and intrinsic indicators of career success, in a sample of 130 graduates who were in the early stages of their careers. Results from hierarchical regression analyses indicated that GMA does not predict any of the success indicators. In contrast, the combination of GMA and three of the Big Five Personality traits, conscientiousness, neuroticism, and openness, is significantly associated with greater early career success and has incremental predictive validity.

The Influence of Gender, Intellectual Ability, Academic Self-Concept, Self-Regulation, Learning Strategies, Popularity and Parent Involvement in Early Adolescence

The present study examined the predictive effects of gender, intellectual ability, self-concept, motivation, learning strategies, popularity and parent involvement on academic achievement. Hiearchical regression analysis were performed with six steps in which each variable was included, among a sample of 1398 high school students (mean age = 12.5; standard deviation = .67) of eight education centers from the province of Alicante (Spain). The results revealed significant predictive effects of all of the variables, explaining 59.1% of the total variance.

Will this time be different for Greece? How to assess its ability to deliver on the reform agenda. CEPS Commentary, 13 March 2015

Taking its inspiration from the ongoing debate on whether this time will be different for Greece and whether Syriza will deliver on its reform promises to the European partners, this Commentary expresses bemusement that the public debate on such an important issue as well as internal discussions among senior policy-makers frequently resort to ‘gut feelings’ or simple stereotypes. To counteract this tendency, the author presents a simple analytical framework that can be used to assess the likelihood that a government will deliver on its reform agenda. Its purpose is not to allow for a precise probabilistic calculation, but to enable better structuring of the knowledge we have. It emphasises that the change depends NOT only on the capacity of the state to design and deliver policies, but even more crucially on state autonomy from both illegitimate and legitimate interests and cognitive models used by policy-makers to make sense of the world.

The effects of age, spatial ability, and navigational information on navigation performance.

Federal Highway Administration, Washington, D.C.

Bicyclists' inclination and ability to search behind before turning left. Interim report.

National Highway Traffic Safety Administration, Washington, D.C.

Imaging ability of collimators in nuclear medicine /

Mode of access: Internet.

Ability for hire.

Mode of access: Internet.

The ability of submerged subjects to sense the gravitational vertical, a pilot study.

"Internal research no. 993-044."

Personal expression; a course of training to develop personality and conversational ability.

Mode of access: Internet.