Anti-CRISPR proteins function through thermodynamic tuning and allosteric regulation of CRISPR RNA-guided surveillance complex.

CRISPR RNA-guided detection and degradation of foreign DNA is a dynamic process. Viruses can interfere with this cellular defense by expressing small proteins called anti-CRISPRs. While structural models of anti-CRISPRs bound to their target complex provide static snapshots that inform mechanism, the dynamics and thermodynamics of these interactions are often overlooked. Here, we use hydrogen deuterium exchange-mass spectrometry (HDX-MS) and differential scanning fluorimetry (DSF) experiments to determine how anti-CRISPR binding impacts the conformational landscape of the type IF CRISPR RNA guided surveillance complex (Csy) upon binding of two different anti-CRISPR proteins (AcrIF9 and AcrIF2). The results demonstrate that AcrIF2 binding relies on enthalpic stabilization, whereas AcrIF9 uses an entropy driven reaction to bind the CRISPR RNA-guided surveillance complex. Collectively, this work reveals the thermodynamic basis and mechanistic versatility of anti-CRISPR-mediated immune suppression. More broadly, this work presents a striking example of how allosteric effectors are employed to regulate nucleoprotein complexes.

How much can you drink before driving? The influence of riding with impaired adults and peers on the driving behaviors of urban and rural youth.

AIMS: Following an ecological model to specify risks for impaired driving, we assessed the effects of youth attitudes about substance use and their experiences of riding in cars with adults and peers who drove after drinking alcohol or smoking cannabis on the youths' own driving after drinking or using cannabis. DESIGN AND METHODS: Participants were 2594 students in grades 10 and 12 (mean age = 16 years and 2 months; 50% girls) from public high schools in urban (994) and rural communities (1600) on Vancouver Island in British Columbia, Canada; 1192 of these were new drivers with restricted licenses. Self-report data were collected in anonymous questionnaires. Regression analyses were used to assess the independent and interacting effects of youth attitudes about substance use and their experiences of riding in cars with adults or peers who drove after drinking alcohol or smoking cannabis on youth driving. FINDINGS: Youth driving risk behaviors were associated independently with their own high-risk attitudes and experiences riding with peers who drink alcohol or use cannabis and drive. However, risks were highest for the youth who also report more frequent experiences of riding with adults who drink alcohol or use cannabis and drive. CONCLUSIONS: Prevention efforts should be expanded to include the adults and peers who are role models for new drivers and to increase youths' awareness of their own responsibilities for their personal safety as passengers.