Biology subjects

Brook, M.

Publications and source records attributed to Brook, M..

5 recordsLinked to original sources

bioRxiv2026

Lysine acetylation facilitates competitive recruitment of translation termination factor eRF3a by PABPC1 C-terminal domain

In eukaryotic cells, almost all mRNA transcripts are polyadenylated in the nucleus, with the poly(A)-tail being critical for their export, translation and stability. PABPC1 is a multifunctional RNA-binding protein (RBP) that binds poly(A)-tails and is a key driver of mRNA post-transcriptional regulation. It achieves this in part by interacting with a broad range of PAM2-motif containing proteins that bind the same common site within PABPC1s PABC domain but result in diverse regulatory outcomes. Regulation of this intricate network of interactions remains poorly understood but our previous studies revealed the regulatory potential of mutually-exclusive acetylation or dimethylation of a key lysine sidechain (K606). To address this, we undertook a systematic study of PAM2-motifs, identifying translation termination factor eRF3a and translation repressor PAIP2 as having a >ten-fold higher affinity for PABPC1 relative to eleven other PAM2-motif partner proteins. Acetylation of PABPC1 at K606 specifically enhances its affinity for eRF3a in vitro, and we present the crystal structure of the specifically K606 acetylated C-terminal domain of PABPC1 in complex with the PAM2 motif of eRF3a, providing structural and functional insights into how acetylation facilitates competitive eRF3a recruitment to PABPC1 and mRNA. We further demonstrate that K606 acetylation promotes PABPC1-eRF3a interaction in cells, thereby antagonising other PAM2-dependent RNA effectors. Taken together, our results show that acetylation of lysine 606 in the PABC domain can specifically modulate PABPC1 partner selectivity, providing crucial insight into how PABPC1 multifunctionality can be regulated to coordinate mRNA usage and fate in the cytoplasm.

molecular biology↗

bioRxiv2025

Mammalian PABPC4 is non-essential, but has roles in growth, post-natal survival and haematopoiesis.

Cytoplasmic poly(A)-binding proteins (PABPCs) are multifunctional RNA-binding proteins which play crucial roles in mRNA translation and stability. In mammals, two family members, PABPC1 and PABPC4 appear widely expressed, but the consequences of their loss of function in vivo remain unknown, despite PABPC4 being implicated in a variety of human diseases. We address this knowledge gap using a series of Pabpc4 knock-out mice. Unexpectedly, we reveal that mammalian PABPC4 is not essential for development, contrary to findings in non-mammalian vertebrates. However, its loss affects birth weight, post-natal growth trajectories and survival, although these were not tightly associated. Growth to adulthood was impacted in a sexually dimorphic manner. Viable Pabpc4-deficient mice allowed us to interrogate roles of Pabpc4 that may affect human health. Anaemia represents a large global health burden, and work in a red blood cell (RBC) model suggested a potential role in haemoglobin synthesis. Surprisingly in vivo, we find that Pabpc4 loss did not reduce haemoglobin levels but caused microcytic RBCs and altered RBC distribution width. Moreover, conditional genetic approaches established that this was not a red blood cell intrinsic effect. Taken together, this work provides unprecedented insights into the in vivo functions of mammalian PABPC4, and caution against inferring mammalian PABPC function from work in cell-based models and/or non-mammalian species. The generated mouse lines also form a valuable resource with which to further investigate the roles of PABPC4 in health and disease. Significance StatementRNA-binding protein (RBP)-mediated post-transcriptional regulation is essential for life and life-long health. Cytoplasmic poly(A)-binding proteins are family of RBPs that regulate multiple aspects of cytoplasmic mRNA fate. PABPC4 is an understudied family member, genetically associated with a wide range of human diseases. By creating a series of Pabpc4 knock-out mice, we surprisingly find it is not essential but is required for normal post-natal survival and growth and normal red blood cell development. Our results emphasise the importance of whole organism studies for understanding mammalian PABP function, uncovering differences between function inferred from other vertebrate species and prior cell-based work. The generated mice also provide a valuable resource for exploring its potential broad roles in human disease.

molecular biology↗

bioRxiv2021

Effects of High-Volume versus High-Load Resistance Training on Skeletal Muscle Growth and Molecular Adaptations

AimWe evaluated the effects of higher-load (HL) versus (lower-load) higher-volume (HV) resistance training on skeletal muscle hypertrophy, strength, and muscle-level molecular markers. MethodsTrained men (n=15, age: 23{+/-}3 y; training experience: 7{+/-}3 y) performed unilateral lower body training for 6 weeks (3x weekly), where single legs were assigned to HV and HL paradigms. Vastus lateralis (VL) biopsies were obtained prior to study initiation (PRE) as well as 3 days (POST) and 10 days following the last bout (POSTPR). Body composition and strength tests were performed at each testing session, and biochemical assays were performed on muscle tissue after study completion. Two-way within subjects repeated measures ANOVAs were performed on all dependent variables except tracer data, which was compared using dependent samples t-tests. ResultsA significant (p<0.05) interaction existed for unilateral leg extension 1RM (HV<HL at POST and POSTPR). Six-week integrated sarcoplasmic protein synthesis (iSarcoPS) rates were higher in the HV versus HL leg, while no difference between legs existed for integrated myofibrillar protein synthesis rates. Main time effects existed for unilateral leg press strength (PRE<POST and POSTPR), knee extensor peak torque (PRE and POST<POSTPR), dual-energy x-ray absorptiometry (DXA)-derived upper leg lean mass (PRE<POST and POSTPR), ultrasound-derived VL thickness (PRE and POSTPR<POST), sarcoplasmic protein concentrations (POST and POSTPR<PRE), and tropomyosin and troponin protein abundances (POST and POSTPR<PRE). ConclusionsWith the exception of differences in leg extensor strength and iSarcoPS between legs, our data suggest that short-term (6 weeks) HV and HL training elicit similar hypertrophic, strength, and molecular-level adaptations.

physiology↗

bioRxiv2018

Shared heritability and functional enrichment across six solid cancers

Quantifying the genetic correlation between cancers can provide important insights into the mechanisms driving cancer etiology. Using genome-wide association study summary statistics across six cancer types based on a total of 296,215 cases and 301,319 controls of European ancestry, we estimate the pair-wise genetic correlations between breast, colorectal, head/neck, lung, ovary and prostate cancer, and between cancers and 38 other diseases. We observed statistically significant genetic correlations between lung and head/neck cancer (rg=0.57, p=4.6x10-8), breast and ovarian cancer (rg=0.24, p=7x10-5), breast and lung cancer (rg=0.18, p=1.5x10-6) and breast and colorectal cancer (rg=0.15, p=1.1x10-4). We also found that multiple cancers are genetically correlated with non-cancer traits including smoking, psychiatric diseases and metabolic characteristics. Functional enrichment analysis revealed a significant excess contribution of conserved and regulatory regions to cancer heritability. Our comprehensive analysis of cross-cancer heritability suggests that solid tumors arising across tissues share in part a common germline genetic basis.

genomics↗

bioRxiv2018

Comparative validation of breast cancer risk prediction models and projections for future risk stratification

BackgroundWell-validated risk models are critical for risk stratified breast cancer prevention. We used the Individualized Coherent Absolute Risk Estimation (iCARE) tool for comparative model validation of five-year risk of invasive breast cancer in a prospective cohort, and to make projections for population risk stratification.\n\nMethodsPerformance of two recently developed models, iCARE-BPC3 and iCARE-Lit, were compared with two established models (BCRAT, IBIS) based on classical risk factors in a UK-based cohort of 64,874 women (863 cases) aged 35-74 years. Risk projections in US White non-Hispanic women aged 50-70 years were made to assess potential improvements in risk stratification by adding mammographic breast density (MD) and polygenic risk score (PRS).\n\nResultsThe best calibrated models were iCARE-Lit (expected to observed number of cases (E/O)=0.98 (95% confidence interval [CI]=0.87 to 1.11)) for women younger than 50 years; and iCARE-BPC3 (E/O=1.00 (0.93 to 1.09)) for women 50 years or older. Risk projections using iCARE-BPC3 indicated classical risk factors can identify ~500,000 women at moderate to high risk (>3% five-year risk). Additional information on MD and a PRS based on 172 variants is expected to increase this to ~3.6 million, and among them, ~155,000 invasive breast cancer cases are expected within five years.\n\nConclusionsiCARE models based on classical risk factors perform similarly or better than BCRAT or IBIS. Addition of MD and PRS can lead to substantial improvements in risk stratification. Independent prospective validation of integrated models is needed prior to clinical evaluation risk stratified breast cancer screening and prevention.

epidemiology↗