Active role of elongation factor G in maintaining the mRNA reading frame during translation.

During translation, the ribosome moves along the mRNA one codon at a time with the help of elongation factor G (EF-G). Spontaneous changes in the translational reading frame are extremely rare, yet how the precise triplet-wise step is maintained is not clear. Here, we show that the ribosome is prone to spontaneous frameshifting on mRNA slippery sequences, whereas EF-G restricts frameshifting. EF-G helps to maintain the mRNA reading frame by guiding the A-site transfer RNA during translocation due to specific interactions with the tip of EF-G domain 4. Furthermore, EF-G accelerates ribosome rearrangements that restore the ribosome's control over the codon-anticodon interaction at the end of the movement. Our data explain how the mRNA reading frame is maintained during translation

Peletização de sementes de milheto

The pearl millet seed is small and its size varies, making sowing more difficult. The pelleting technique increases and homogenizes seed size, but it is essential to determine the physical and physiological characteristics of pelleted seeds. The physiological analysis consisted of: first germination count, final germination, speed emergence index, and seedling emergence. Physical analysis consisted of determining the 1000-seed weight, 1000-seed volume and fragmentation. The control treatment did not receive any coating, and the other 36 treatments combined four binders: bentonite, polyvinyl acetate (PVA), polyvinylpyrrolidone (PVP) and methyl cellulose (Methocel®), and nine powder coating products: microcellulose, plaster, vermiculite, magnesium thermophosphate (Yoorin®), phytic acid, dicalcium phosphate, super simple phosphate (SS), monoamonic phosphate (MAP) and reactive phosphate. Among the materials used to form the pearl millet pellet, the most efficient binders were the polyvinyl acetate and the methyl cellulose, and as coaters, the vermiculite and the microcellulose.As sementes de milheto são pequenas e têm tamanho variado, vindo a dificultar a semeadura. A técnica de peletização aumenta e uniformiza a forma das sementes. No entanto, é necessário determinar as características físicas e fisiológicas das sementes peletizadas. Logo, as análises fisiológicas consistiram do teste de germinação, primeira contagem de germinação, índice de velocidade de emergência e emergência final. As análises físicas consistiram do teste de peso de mil sementes, volume de mil sementes e fragmentação. A testemunha foi o tratamento de sementes sem peletização, os demais 36 tratamentos combinaram quatro adesivos: bentonita, acetato de polvinilina (PVA), polivinilpirrolidona (PVP) e metil celulose (Methocel®), e nove produtos de enchimento em pó: microcelulose, gesso, vermiculita, termofosfato magnesiano (Yoorin®), fitina, fosfato bicálcico, fosfato super simples (SS), fosfato monoamônico (MAP) e fosfato natural reativo. Dentre os materiais testados para formar a pelota de semente de milheto nesta pesquisa, os adesivos mais eficientes foram o acetato de polvinilina e a metil celulose. Já os materiais de enchimento em pó mais eficientes foram a vermiculita e a microcelulose.CNP

Comparative Transcriptomic Analysis Identifies a Range of Immunologically Related Functional Elaborations of Lymph Node Associated Lymphatic and Blood Endothelial Cells

Lymphatic and blood vessels are formed by specialized lymphatic endothelial cells (LEC) and blood endothelial cells (BEC), respectively. These endothelial populations not only form peripheral tissue vessels, but also critical supporting structures in secondary lymphoid organs, particularly the lymph node (LN). Lymph node LEC (LN-LEC) also have been shown to have important immunological functions that are not observed in LEC from tissue lymphatics. LN-LEC can maintain peripheral tolerance through direct presentation of self-antigen via MHC-I, leading to CD8 T cell deletion; and through transfer of self-antigen to dendritic cells for presentation via MHC-II, resulting in CD4 T cell anergy. LN-LEC also can capture and archive foreign antigens, transferring them to dendritic cells for maintenance of memory CD8 T cells. The molecular basis for these functional elaborations in LN-LEC remain largely unexplored, and it is also unclear whether blood endothelial cells in LN (LN-BEC) might express similar enhanced immunologic functionality. Here, we used RNA-Seq to compare the transcriptomic profiles of freshly isolated murine LEC and BEC from LN with one another and with freshly isolated LEC from the periphery (diaphragm). We show that LN-LEC, LN-BEC, and diaphragm LEC (D-LEC) are transcriptionally distinct from one another, demonstrating both lineage and tissue-specific functional specializations. Surprisingly, tissue microenvironment differences in gene expression profiles were more numerous than those determined by endothelial cell lineage specification. In this regard, both LN-localized endothelial cell populations show a variety of functional elaborations that suggest how they may function as antigen presenting cells, and also point to as yet unexplored roles in both positive and negative regulation of innate and adaptive immune responses. The present work has defined in depth gene expression differences that point to functional specializations of endothelial cell populations in different anatomical locations, but especially the LN. Beyond the analyses provided here, these data are a resource for future work to uncover mechanisms of endothelial cell functionality

Ileal immune tonus is a prognosis marker of proximal colon cancer in mice and patients

Ileal epithelial cell apoptosis and the local microbiota modulate the effects of oxaliplatin against proximal colon cancer by modulating tumor immunosurveillance. Here, we identified an ileal immune profile associated with the prognosis of colon cancer and responses to chemotherapy. The whole immune ileal transcriptome was upregulated in poor-prognosis patients with proximal colon cancer, while the colonic immunity of healthy and neoplastic areas was downregulated (except for the Th17 fingerprint) in such patients. Similar observations were made across experimental models of implanted and spontaneous murine colon cancer, showing a relationship between carcinogenesis and ileal inflammation. Conversely, oxaliplatin-based chemotherapy could restore a favorable, attenuated ileal immune fingerprint in responders. These results suggest that chemotherapy inversely shapes the immune profile of the ileum-tumor axis, influencing clinical outcome

Development of corn seedlings after a period of exposition to various water potential

Fungicide treated and untreated corn seeds were sets to germinate under varying water availability conditions (0 to -12atm water potential). The seedlings obtained on each individual treatment were subsequently grown under ideal available water conditions. The results permitted to conclude that the occurrence of a water deficit during the on set of the germination process results in reductions on the length of the embryonic structures, particularly the epicotil. This effect attenuates with times due to the increase in growth velocity in environments where water potential is lower. Fungicid treated seeds, did not appear to be consistently vantageous in environments with water stress.Submetendo sementes de milho com e sem tratamento fungicida, a ambientes de germinação variáveis quanto à disponibilidade de água (0 a -12atm), o presente trabalho avaliou o desenvolvimento posterior das plântulas postas em ambientes sem limitações hídricas. Os resultados obtidos permitiram concluir que a ocorrência de déficit hídrico, durante o início do processo de germinação, promove reduções posteriores no comprimento das estruturas embrionárias e, de forma mais acentuada, do epicótilo; apesar disso, esse efeito se atenua com o passar do tempo pela elevação progressiva na velocidade de crescimento em ambientes que oferecem menores potenciais hídricos. Paralelamente, o tratamento fungicida das sementes pode não trazer vantagens em ambientes hidricamente deficientes; esta afirmação, contudo, considera o papel toalha como meio fornecedor de água o que, em contrapartida, exige cautela na sua extrapolação para as condições proporcionadas pelo solo

Structural Basis for Polyproline-Mediated Ribosome Stalling and Rescue by the Translation Elongation Factor EF-P

Ribosomes synthesizing proteins containing consecutive proline residues become stalled and require rescue via the action of uniquely modified translation elongation factors, EF-P in bacteria, or archaeal/eukaryotic a/eIF5A. To date, no structures exist of EF-P or eIF5A in complex with translating ribosomes stalled at polyproline stretches, and thus structural insight into how EF-P/eIF5A rescue these arrested ribosomes has been lacking. Here we present cryo-EM structures of ribosomes stalled on proline stretches, without and with modified EF-P. The structures suggest that the favored conformation of the polyproline-containing nascent chain is incompatible with the peptide exit tunnel of the ribosome and leads to destabilization of the peptidyltRNA. Binding of EF-P stabilizes the P-site tRNA, particularly via interactions between its modification and the CCA end, thereby enforcing an alternative conformation of the polyproline-containing nascent chain, which allows a favorable substrate geometry for peptide bond formation