Smith, N. W. | bioRxiv Science

bioRxiv2024

Evaluation of protein intake and protein quality in New Zealand vegans

Dietary protein provides indispensable amino acids (IAAs) that the body cannot synthesise. Past assessments of total protein intake from vegan populations in western, developed countries were found to be low but not necessarily below daily requirements. However, plant-sourced proteins generally have lower quantities of digestible IAAs as compared to animal-sourced proteins. Simply accounting for protein intake without considering AA profile and digestibility could overestimate protein adequacy among vegans. This study quantified protein intake and quality, as compared to reference intake values among 193 NZ vegans using a four-day food diary. Protein and IAA composition of all foods were derived from New Zealand FoodFiles and the United States Department of Agriculture and adjusted for True Ileal Digestibility (TID). Mean protein intakes for males and females were 0.99 and 0.81 g/kg /day, respectively with 78.8% of males and 74.5% of females meeting the Estimated Average Requirement for protein. Plant-sourced proteins provided 52.9 mg of leucine and 35.7 mg of lysine per gram of protein and were below the reference scoring patterns (leucine: 59mg/g, lysine: 45mg/g). When adjusted to individual body weight, average IAA intakes were above daily requirements, but lysine just met requirements at 31.2 mg/kg of body weight/day (reference: 30 mg/kg/day). Upon TID adjustment, the percentage of vegans meeting adequacy for protein and IAA decreased and only approximately 50% of the cohort could meet lysine and leucine requirements. Hence, lysine and leucine were the most limiting IAAs in the vegan cohorts diet. Legumes and pulses contributed most to overall protein and lysine intake. An increased proportion of legumes and pulses can potentially increase these intakes but must be considered in the context of the whole diet. AA composition and digestibility are important aspects of protein quality when assessing protein adequacy and is of particular importance in restrictive diets.

physiology↗

bioRxiv2018

Werner helicase control of human papillomavirus 16 E1-E2 DNA replication is regulated by SIRT1 deacetylation

Human papillomaviruses (HPV) are double stranded DNA viruses causative in a host of human diseases including several cancers. Following infection two viral proteins, E1 and E2, activate viral replication in association with cellular factors, and stimulate the DNA damage response (DDR) during the replication process. E1-E2 uses homologous replication (HR) to facilitate DNA replication, but an understanding of host factors involved in this process remains incomplete. Previously we demonstrated that the class III deacetylase SIRT1, which can regulate HR, is recruited to E1-E2 replicating DNA and regulates the level of replication. Here we demonstrate that SIRT1 promotes the fidelity of E1-E2 replication and that the absence of SIRT1 results in reduced recruitment of the DNA repair protein Werner helicase (WRN) to E1-E2 replicating DNA. CRISPR/Cas9 editing demonstrates that WRN, like SIRT1, regulates the quantity and fidelity of E1-E2 replication. This is the first report of WRN regulation of E1-E2 DNA replication, or a role for WRN in the HPV life cycle. In the absence of SIRT1 there is an increased acetylation and stability of WRN, but a reduced ability to interact with E1-E2 replicating DNA. We present a model in which E1-E2 replication turns on the DDR stimulating SIRT1 deacetylation of WRN. This deacetylation promotes WRN interaction with E1-E2 replicating DNA to control the quantity and fidelity of replication. As well as offering a crucial insight into HPV replication control, this system offers a unique model for investigating the link between SIRT1 and WRN in controlling replication in mammalian cells.\n\nImportanceHPV16 is the major viral human carcinogen, responsible for between 3 and 4% of all cancers worldwide. Following infection this virus activates the DNA damage response (DDR) to promote its life cycle, and recruits DDR proteins to its replicating DNA in order to facilitate homologous recombination during replication. This promotes the production of viable viral progeny. Our understanding of how HPV16 replication interacts with the DDR remains incomplete. Here we demonstrate that the cellular deacetylase SIRT1, which is a part of the E1-E2 replication complex, regulates recruitment of the DNA repair protein WRN to the replicating DNA. We demonstrate that WRN regulates the level and fidelity of E1-E2 replication. Overall the results suggest a mechanism where SIRT1 deacetylation of WRN promotes its interaction with E1-E2 replicating DNA to control the levels and fidelity of that replication.

microbiology↗