UBR1 promotes protein kinase quality control and sensitizes cells to Hsp90 inhibition.

UBR1 and UBR2 are N-recognin ubiquitin ligases that function in the N-end rule degradation pathway. In yeast, the UBR1 homologue also functions by N-end rule independent means to promote degradation of misfolded proteins generated by treatment of cells with geldanamycin, a small molecule inhibitor of Hsp90. Based on these studies we examined the role of mammalian UBR1 and UBR2 in the degradation of protein kinase clients upon Hsp90 inhibition. Our findings show that protein kinase clients Akt and Cdk4 are still degraded in mouse Ubr1(-)/(-) cells treated with geldanamycin, but that their levels recover much more rapidly than is found in wild type cells. These findings correlate with increased induction of Hsp90 expression in the Ubr1(-)/(-) cells compared with wild type cells. We also observed a reduction of UBR1 protein levels in geldanamycin-treated mouse embryonic fibroblasts and human breast cancer cells, suggesting that UBR1 is an Hsp90 client. Further studies revealed a functional overlap between UBR1 and the quality control ubiquitin ligase, CHIP. Our findings show that UBR1 function is conserved in controlling the levels of Hsp90-dependent protein kinases upon geldanamycin treatment, and suggest that it plays a role in determining the sensitivity of cancer cells to the chemotherapeutic effects of Hsp90 inhibitors.

Role of Metals on SARS-CoV-2 Infection: a Review of Recent Epidemiological Studies.

PURPOSE OF REVIEW: Metals and metalloids are known for their nutritional as well as toxic effects in humans. In the context of the SARS-CoV-2 pandemic, understanding the role of metals on COVID-19 infection is becoming important due to their role in infectious diseases. During the past 2 years, a significant number of studies have examined the impact of metals and metalloids on COVID-19 morbidity and mortality. We conducted a systematic review of peer-reviewed manuscripts on the association of metals and metalloids with SARS-CoV-2 infection and COVID-19 severity published since the onset of the pandemic. RECENT FINDINGS: We searched for epidemiological studies available through the PubMed database published from January 2020 to December 2022. Of 92 studies identified, 20 met our inclusion criteria. These articles investigated the association of zinc (Zn), iron (Fe), selenium (Se), manganese (Mn), cadmium (Cd), arsenic (As), copper (Cu), magnesium (Mg), chromium (Cr), and/or lead (Pb) levels on SARS-CoV-2 infection and/or COVID-19 severity. Of the ten metals and metalloids of interest that reported either positive, negative, or no associations, Zn yielded the highest number of articles (n = 13), followed by epidemiological studies on Se (n = 7) and Fe (n = 5). Elevated serum Zn and Se were associated with reduced COVID-19 severity and mortality. Similarly, higher levels of serum Fe were associated with lower levels of cellular damage and symptoms of SARS-CoV-2 infection and with faster recovery from COVID-19. On the other hand, higher serum and urinary Cu and serum Mg levels were associated with higher COVID-19 severity and mortality. Along with the positive or negative effects, some studies reported no impact of metals on SARS-CoV-2 infection. This systematic review suggests that metals, particularly Zn, Fe, and Se, may help reduce the severity of COVID-19, while Cu and Mg may aggravate it. Our review suggests that future pandemic mitigation strategies may evaluate the role of Zn, Se, and Fe as potential therapeutic interventions.

In vitro antibacterial activity of cefiderocol against recent multidrug-resistant carbapenem-nonsusceptible Enterobacterales isolates.

Cefiderocol, a siderophore-containing cephalosporin with broad-spectrum antimicrobial activity against many ß-lactam-resistant Gram-negative bacteria, was tested by broth microdilution against 104 carbapenem-non-susceptible Enterobacterales clinical isolates from 2011 to 2018. Carbapenemase identification was determined using PCR followed by targeted gene sequencing or whole genome sequencing (WGS). All isolates were multidrug-resistant, 89.4% (93/104) and produced a serine (KPC or SME) carbapenemase, with as many as four ß-lactamases present. A VIM-1 or NDM-1 metallo-ß-lactamase was confirmed in 6.7% of the isolates (N = 5 and 2, respectively). All isolates were susceptible to cefiderocol, unlike the comparator agents. Susceptibility for comparators ranged from 24.0% for meropenem to 91.3%, 92.3% and 96.1% for imipenem-relebactam, ceftazidime-avibactam and meropenem-vaborbactam, respectively; 48.1%, 75.2% and 79.8% of the isolates were susceptible to omadacycline, colistin and eravacycline, respectively. Two isolates with cefiderocol MICs of 2 mg/L had mutations or deletions of the iron transport genes fhuA/E or fepA, as determined by WGS.

Association between Handwashing Behavior and Infectious Diseases among Low-Income Community Children in Urban New Delhi, India: A Cross-Sectional Study.

Diarrheal diseases and respiratory infections (RI) are two leading causes of childhood mortality in low and middle-income countries. Effective handwashing at critical time-points may mitigate these diseases. However, there is a lack of published data investigating this association in school-aged children in India. This study is part of a larger prospective handwashing intervention study in a low-income community in New Delhi, India examining the associations between handwashing behavior and diarrhea and RI in schoolchildren. This current study reports the findings of the baseline survey administered to 272 mother-child dyads. Children aged 8-12 years, and their mothers, were recruited from six schools. A baseline questionnaire was used to collect sociodemographic data, handwash behavior, and mother-reported recent diarrhea and RI incidence among the children. Handwashing before and after preparing food, after defecation, and after cleaning dishes significantly reduced the odds of diarrhea by over 70%, and of RI by over 56%. Using a clean cloth after handwashing lowered odds of diarrhea and RI by 72% and 63% respectively. Around 60% of the participants believed that handwashing could prevent diarrhea and RI in their children. There was a low prevalence of handwashing at critical time-points and a poor perception regarding handwashing benefits. To improve handwashing behavior, hygiene promotion programs need to understand what motivates and hinders handwashing in vulnerable populations.

Typing of human rotaviruses: nucleotide mismatches between the VP7 gene and primer are associated with genotyping failure.

BACKGROUND: Rotavirus genotyping is performed by using reverse transcription PCR with type-specific-primers. Because the high rotavirus mutation rate generates an extensive genomic variation, different G-type-specific primer sets are applied in different geographical locations. In Bangladesh, a significant proportion (36.9%) of the rotavirus strains isolated in 2002 could not be G-typed using the routinely used primer set. To investigate the reason why the strains were untypeable, nucleotide sequencing of the VP7 genes was performed. RESULTS: Four nucleotide substitutions at the G1 primer-binding site of the VP7 gene of Bangladeshi G1 rotaviruses rendered a major proportion of circulating strains untypeable using the routine primer set. Using an alternative primer set, we could identify G1 rotaviruses as the most prevalent genotype (44.8%), followed by G9 (21.7%), G2 (15.0%) and G4 (13.8%). CONCLUSION: Because of the natural variation in the rotaviral gene sequences, close monitoring of rotavirus genotyping methods is important.

Specificity in the actions of the UBR1 ubiquitin ligase in the degradation of nuclear receptors.

The UBR1 ubiquitin ligase promotes degradation of proteins via the N-end rule and by another mechanism that detects a misfolded conformation. Although UBR1 was shown recently to act on protein kinases whose misfolding was promoted by inhibition of Hsp90, it was unknown whether this ubiquitin ligase targeted other client types of the chaperone. We analyzed the role of UBR1 in the degradation of nuclear receptors that are classical clients of Hsp90. Our results showed that UBR1 deletion results in impaired degradation of the glucocorticoid receptor and the androgen receptor but not the estrogen receptor α. These findings demonstrate specificity in the actions of the UBR1 ubiquitin ligase in the degradation of Hsp90 clients in the presence of small molecule inhibitors that promote client misfolding.

Ubr1 and Ubr2 function in a quality control pathway for degradation of unfolded cytosolic proteins.

Quality control systems facilitate polypeptide folding and degradation to maintain protein homeostasis. Molecular chaperones promote folding, whereas the ubiquitin/proteasome system mediates degradation. We show here that Saccharomyces cerevisiae Ubr1 and Ubr2 ubiquitin ligases promote degradation of unfolded or misfolded cytosolic polypeptides. Ubr1 also catalyzes ubiquitinylation of denatured but not native luciferase in a purified system. This activity is based on the direct interaction of denatured luciferase with Ubr1, although Hsp70 stimulates polyubiquitinylation of the denatured substrate. We also report that loss of Ubr1 and Ubr2 function suppressed the growth arrest phenotype resulting from chaperone mutation. This correlates with increased protein kinase maturation and indicates partitioning of foldable conformers toward the proteasome. Our findings, based on the efficiency of this quality control system, suggest that the cell trades growth potential to avert the potential toxicity associated with accumulation of unfolded or misfolded proteins. Ubr1 and Ubr2 therefore represent E3 components of a novel quality control pathway for proteins synthesized on cytosolic ribosomes.

Prevalence of G2P[4] and G12P[6] rotavirus, Bangladesh.

Approximately 20,000 stool specimens from patients with diarrhea visiting 1 urban and 1 rural hospital in Bangladesh during January 2001-May 2006 were tested for group A rotavirus antigen, and 4,712 (24.0%) were positive. G and P genotyping was performed on a subset of 10% of the positive samples (n = 471). During the 2001-2005 rotavirus seasons, G1P[8] (36.4%) and G9P[8] (27.7%) were the dominant strains, but G2[4] and G12P[6] were present in 15.4% and 3.1% of the rotavirus-positive patients, respectively. During the 2005-06 rotavirus season, G2P[4] (43.2%) appeared as the most prevalent strain, and G12P[6] became a more prevalent strain (11.1%) during this season. Because recently licensed rotavirus vaccines include only the P[8] specificity, it is unknown how the vaccines will perform in settings where non-P[8] types are prevalent.