Cross-Species Transmission of Bat Coronaviruses in the Americas: Contrasting Patterns between Alphacoronavirus and Betacoronavirus

Bats harbor the largest number of coronavirus (CoV) species among mammals, serving as major reservoirs of alphaCoVs and betaCoVs, which can jump between bat species or to different mammalian hosts, including humans. Bat-CoV diversity is correlated with host taxonomic diversity, with the highest number of CoV species found in areas with the highest levels of bat species richness. Although the Americas harbor a unique and distinctive CoV diversity, no cross-species transmission (CST) or phylogeographic analysis has yet been performed. This study analyzes a large sequence data set from across the Americas through a Bayesian framework to understand how codivergence and cross-species transmission have shaped long-term bat-CoV evolution and ultimately identify bat hosts and regions where the risk of CST is the highest. Substantial levels of CST were found only among alphaCoVs. In contrast, cospeciation prevailed along the evolution of betaCoVs. Brazil is the center of diversification for both alpha and betaCoVs, with the highest levels of bat species richness. The bat family Phyllostomidae has played a key role in the evolution of American bat-CoVs, supported by the highest values of host transition rates. Although the conclusions drawn from this study are supported by biological/ecological evidence, it is likely that novel lineages will be discovered, which could also reveal undetected CSTs given that sequences are available from 11 of the 35 countries encompassing the Americas. The findings of this study can be useful for conducting targeted discovery of bat-CoVs in the region, especially in countries of the Americas with no reported sequences.Fil: Caraballo, Diego Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentin

Spatial and temporal divergence of the torquatus species group of the subterranean rodent Ctenomys

Subterranean rodents of the genus Ctenomys have experienced an explosive radiation and rapidly colonized the southern cone of South America. The torquatus group, one of the main groups of the genus, comprises several species and species complexes which inhabit the eastern part of the distribution of Ctenomys including southern Brazil, northern and central Uruguay and north-eastern Argentina. This group has undergone a high chromosomal diversification with diploid numbers varying from 41 to 70. The aim of this study was to investigate the origins of the torquatus group as well as its diversification patterns in relation to geography and cladogenesis. Based on mitochondrial cytochrome b nucleotide sequences we conducted a Bayesian multi-calibrated relaxed clock analysis to estimate the ages of the torquatus group and its main lineages. Using the estimated evolutionary rate we performed a continuous phylogeographic analysis, using a relaxed random walk model to reconstruct the geographic diffusion of the torquatus group in a temporal frame. The torquatus group originated during the early Pleistocene between 1.25 and 2.32 million years from the present in a region that includes the northwest of Uruguay and the southeast of the Brazilian state of Río Grande do Sul. Most lineages have dispersed early towards their present distribution areas going through subsequent range expansions in the last 800,000 – 700,000 years. Ctenomys torquatus went through a rapid range expansion for the last 200,000 years, becoming the most widespread species of the group. The colonization of the Corrientes and Entre Ríos Argentinean provinces supposes at least two crossing events across the Uruguay River between 1.0 and 0.5 million years before the present, in the context of a cold and dry paleoenvironment. The resulting temporal and geographic frame enables the comprehension of the incidence of both, the amplitude of distribution areas and divergence times into the patterns of chromosomal diversification found in the group.Fil: Caraballo, Diego Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Rossi, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

Origin and Evolution of Two Independently Duplicated Genes Encoding UDP- Glucose: Glycoprotein Glucosyltransferases in Caenorhabditis and Vertebrates

UDP- glucose: glycoprotein glucosyltransferase (UGGT) is a protein that operates as the gatekeeper for the endoplasmic reticulum (ER) quality control mechanism of glycoprotein folding. It is known that vertebrates and Caenorhabditis genomes harbor two uggt gene copies that exhibit differences in their properties.Bayesian phylogenetic inference based on 195 UGGT and UGGT-like protein sequences of an ample spectrum of eukaryotic species showed that uggt genes went through independent duplications in Caenorhabditis and vertebrates. In both lineages, the catalytic domain of the duplicated genes was subjected to a strong purifying selective pressure, while the recognition domain was subjected to episodic positive diversifying selection. Selective relaxation in the recognition domain was more pronounced in Caenorhabditis uggt-b than in vertebrates uggt-2. Structural bioinformatics analysis revealed that Caenorhabditis UGGT-b protein lacks essential sequences proposed to be involved in the recognition of unfolded proteins. When we assayed glucosyltrasferase activity of a chimeric protein composed by Caenorhabditis uggt-b recognition domain fused to S. pombe catalytic domain expressed in yeast, no activity was detected.The present results support the conservation of the UGGT activity in the catalytic domain and a putative divergent function of the recognition domain for the UGGT2 protein in vertebrates, which would have gone through a specialization process. In Caenorhabditis, uggt-b evolved under different constraints compared to uggt-a which, by means of a putative neofunctionalization process, resulted in a non-redundant paralog. The non-canonical function of uggt-b in the worm lineage highlights the need to take precautions before generalizing gene functions in model organisms.Fil: Caraballo, Diego Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Buzzi, Lucila Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Acosta Montalvo, Ana Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Modenutti, Carlos Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Castro, Olga Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Rossi, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

A novel terrestrial rabies virus lineage occurring in south america: Origin, diversification, and evidence of contact between wild and domestic cycles

The rabies virus (RABV) is characterized by a history dominated by host shifts within and among bats and carnivores. One of the main outcomes of long-term RABV maintenance in dogs was the establishment of variants in a wide variety of mesocarnivores. In this study, we present the most comprehensive phylogenetic and phylogeographic analysis, contributing to a better understanding of the origins, diversification, and the role of different host species in the evolution and diffusion of a dog-related variant endemic of South America. A total of 237 complete Nucleoprotein gene sequences were studied, corresponding to wild and domestic species, performing selection analyses, ancestral states reconstructions, and recombination analyses. This variant originated in Brazil and disseminated through Argentina and Paraguay, where a previously unknown lineage was found. A single host shift was identified in the phylogeny, from dog to the crab-eating fox (Cerdocyon thous) in the Northeast of Brazil. Although this process occurred in a background of purifying selection, there is evidence of adaptive evolution-or selection of sub-consensus sequences-in internal branches after the host shift. The interaction of domestic and wild cycles persisted after host switching, as revealed by spillover and putative recombination events.Fil: Caraballo, Diego Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Lema, Cristina. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio E Instituto de Salud "Dr. C. G. Malbran". Departamento Virus; ArgentinaFil: Novaro, Laura. Ministerio de Agricultura, Ganadería, Pesca y Alimento. Servicio Nacional de Sanidad y Calidad Agroalimentaria; ArgentinaFil: Gury Dohmen, Federico. Gobierno de la Ciudad Autónoma de Buenos Aires. Ministerio de Salud. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Russo, Susana. Ministerio de Agricultura, Ganadería, Pesca y Alimento. Servicio Nacional de Sanidad y Calidad Agroalimentaria; ArgentinaFil: Beltrán, Fernando J.. Gobierno de la Ciudad Autónoma de Buenos Aires. Ministerio de Salud. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Palacios, Gustavo. Icahn School of Medicine at Mount Sinai; Estados UnidosFil: Cisterna, Daniel Marcelo. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio E Instituto de Salud "Dr. C. G. Malbran". Departamento Virus; Argentin

TSSK3, a novel target for male contraception, is required for spermiogenesis

We have previously shown that members of the family of testis-specific serine/threonine kinases (TSSKs) are post-meiotically expressed in testicular germ cells and in mature sperm in mammals. The restricted post-meiotic expression of TSSKs as well as the importance of phosphorylation in signaling processes strongly suggest that TSSKs have an important role in germ cell differentiation and/or sperm function. This prediction has been supported by the reported sterile phenotype of the TSSK6 knock-out (KO) mice and of the double TSSK1/TSSK2 KO. The aim of this study was to develop KO mouse models of TSSK3 and to validate this kinase as a target for the development of a male contraceptive. We used CRISPR/Cas9 technology to generate the TSSK3 KO allele on B6D2F1 background mice. Male heterozygous pups were used to establish three independent TSSK3 KO lines. After natural mating of TSSK3 KO males, females that presented a plug (indicative of mating) were monitored for the following 24 days and no pregnancies or pups were found. Sperm numbers were drastically reduced in all three KO lines and, remarkably, round spermatids were detected in the cauda epididymis of KO mice. From the small population of sperm recovered, severe morphology defects were detected. Our results indicate an essential role of TSSK3 in spermiogenesis and support this kinase as a suitable candidate for the development of novel nonhormonal male contraceptives.Fil: Nayyab, Saman. University of Massachussets; Estados UnidosFil: Gervasi, María G.. University of Massachussets; Estados UnidosFil: Tourzani, Darya A.. University of Massachussets; Estados UnidosFil: Caraballo, Diego Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Jha, Kula N.. No especifíca;Fil: Teves, Maria E.. University of Virginia; Estados UnidosFil: Cui, Wei. University of Massachussets; Estados UnidosFil: Georg, Gunda I.. University of Minnesota; Estados UnidosFil: Visconti, Pablo E.. University of Massachussets; Estados UnidosFil: Salicioni, Ana M.. University of Massachussets; Estados Unido

Evolution of a species complex of the genus Ctenomys (Octodontidae, Rodentia) from northeastern Argentina: phylogeny, chromosomal variability and satellite DNA dynamics

Los tuco-tucos, roedores subterráneos del género Ctenomys, de la provincia de Corrientes habitan en gran parte de su área de distribución en ambientes inestables tanto desde el punto de vista espacial como temporal. Constituyen un excelente modelo para estudiar la especiación y la hibridación, y el rol de la evolución cromosómica en estos procesos, un rasgo conspicuo en este grupo. Unas 39 poblaciones fueron descriptas en este grupo, la mayoría de status taxonómico indefinido. La variabilidad en el número diploide (2n) y número fundamental (NF) es inusualmente elevada en este grupo (2n=41-70, NF=76-84). En este trabajo se obtuvo una filogenia molecular que incluye representantes de 23 poblaciones correntinas, utilizando los marcadores mitocondriales citocromo b, citocromo oxidasa I y región control (D-loop). El grupo Corrientes resultó monofilético. Las especies previamente descriptas C. perrensi y C. dorbignyi no resultaron monofiléticas. Se propone el subgrupo iberá como linaje evolutivo diferenciado e independiente. Por otro lado, se obtuvieron cariomorfos de 33 individuos. El cariomorfo 2n=70 NF=84 ocurre en dos linajes basales en el grupo Corrientes, y en la especie hermana C. pearsoni, sería ancestral y habría sufrido reducciones en 2n y NF vía fusiones céntricas y en tándem, principalmente. Se exploró la relación entre la dinámica del principal ADN satélite de los tucos (SRPC) y la variabilidad cromosómica en Corrientes. Se analizó también la variación intra/inter poblacional del número de copias y de la secuencia del satélite SRPC. El satélite SRPC siguió un patrón conservativo en algunos linajes pero altamente dinámico en otros. La evolución de la secuencia y el número de copias de SRPC es compatible con la hipótesis de una biblioteca ancestral, cuyas variantes están presentes en todos los linajes, aunque en proporciones diferentes

Evolución de un complejo de especies de Ctenomys (Octodontidae, Rodentia) del noreste argentino: filogenia, variabilidad cromosómica y dinámica del ADN satélite

Los tuco-tucos, roedores subterráneos del género Ctenomys, de la provincia de Corrientes habitan en gran parte de su área de distribución en ambientes inestables tanto desde el punto de vista espacial como temporal. Constituyen un excelente modelo para estudiar la especiación y la hibridación, y el rol de la evolución cromosómica en estos procesos, un rasgo conspicuo en este grupo. Unas 39 poblaciones fueron descriptas en este grupo, la mayoría de status taxonómico indefinido. La variabilidad en el número diploide (2n) y número fundamental (NF) es inusualmente elevada en este grupo (2n=41-70, NF=76-84). En este trabajo se obtuvo una filogenia molecular que incluye representantes de 23 poblaciones correntinas, utilizando los marcadores mitocondriales citocromo b, citocromo oxidasa I y región control (D-loop). El grupo Corrientes resultó monofilético. Las especies previamente descriptas C. perrensi y C. dorbignyi no resultaron monofiléticas. Se propone el subgrupo iberá como linaje evolutivo diferenciado e independiente. Por otro lado, se obtuvieron cariomorfos de 33 individuos. El cariomorfo 2n=70 NF=84 ocurre en dos linajes basales en el grupo Corrientes, y en la especie hermana C. pearsoni, sería ancestral y habría sufrido reducciones en 2n y NF vía fusiones céntricas y en tándem, principalmente. Se exploró la relación entre la dinámica del principal ADN satélite de los tucos (SRPC) y la variabilidad cromosómica en Corrientes. Se analizó también la variación intra/inter poblacional del número de copias y de la secuencia del satélite SRPC. El satélite SRPC siguió un patrón conservativo en algunos linajes pero altamente dinámico en otros. La evolución de la secuencia y el número de copias de SRPC es compatible con la hipótesis de una biblioteca ancestral, cuyas variantes están presentes en todos los linajes, aunque en proporciones diferentes

Integrative lineage delimitation in rodents of the Ctenomys Corrientes group

The tuco-tucos rodents (genus Ctenomys) of the Corrientes group comprise several populations that inhabit the vast area under the influence of the Iberá wetland. Lineage delimitation within the recently diverged Corrientes group is a challenging task as morphological differentiation is not conspicuous between populations. However, delimitation is crucial for evolutionary studies and conservation issues. In this study, we performed a phylogenetic analysis including cytochrome b (cyt-b) sequences from taxa that had never been studied in a comprehensive context. We integrated previously published chromosomal studies, mitochondrial phylogenies and simple sequence repeat (SSR) variability analyses, and applied a delimitation criterion over the basis of chromosomal incompatibilities and genetic exclusivity. Under this integrative approach seven independently evolving lineages were delimited in the Corrientes group: Ctenomys roigi, which conserves its former definition, Ctenomys dorbignyi and Ctenomys perrensi complex which were redefined, Sarandicito which includes the population of Paraje Sarandicito and probably a group of nearby poorly studied populations, and Iberá i, ii and iii distributed at both sides of the Iberá wetland. We discuss future perspectives to evaluate the proposed lineages and conservation issues concerning these tuco-tucos.Fil: Caraballo, Diego Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Rossi, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

Diversity of tuco-tucos (Ctenomys, Rodentia) in the Northeastern wetlands from Argentina: mitochondrial phylogeny and chromosomal evolution

Tuco-tucos (small subterranean rodents of the genus Ctenomys) that inhabit sandy soils of the area under the influence of the second largest wetland of South America, in Northeastern Argentina (Corrientes province), are a complex of species and forms whose taxonomic status were not defined, nor are the evolutionary relationships among them. The tuco-tuco populations of this area exhibit one of the most ample grades of chromosomal variability within the genus. In order to analyze evolutionary relationships within the Corrientes group and its chromosomal variability, we completed the missing karyotypic information and performed a phylogenetic analysis. We obtained partial sequences of three mitochondrial markers: D-loop, cytochrome b and cytochrome oxidase I. The Corrientes group was monophyletic and split into three main clades that grouped related karyomorphs. The phylogeny suggested an ancestral condition of the karyomorph with diploid number (2n) 70 and fundamental number (FN) 84 that has evolved mainly via reductions of the FN although amplifications occurred in certain lineages. We discuss the relationship between patterns of chromosomal variability and species and groups boundaries. From the three main clades the one named ibera´ exhibited a remarkable karyotypic homogeneity, and could be considered as an independent and cohesive evolutionary lineage. On the contrary, the former recognized species C. dorbignyi is a polyphyletic lineage and hence its systematic classification should be reviewed.Fil: Caraballo, Diego Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Abruzzese, Giselle Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Rossi, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

The library model for satellite DNA evolution: a case study with the rodents of the genus Ctenomys (Octodontidae) from the Iberá marsh, Argentina

On the basement of the library model of satellite DNA evolution is the differential amplification of subfamilies through lineages diversification. However, this idea has rarely been explored from an experimental point of view. In the present work, we analyzed copy number and sequence variability of RPCS (repetitive PvuII Ctenomys sequence), the major satellite DNA present in the genomes of the rodents of the genus Ctenomys, in a closely related group of species and forms inhabiting the Iberá marsh in Argentina. We studied the dependence of these two parameters at the intrapopulation level because in the case of interbreeding genomes, differences in RPCS copy number are due to recent amplification/contraction events. We found an inverse relationship among RPCS copy number and sequence variability: amplifications lead to a decrease in sequence variability, by means of biased homogenization of the overall satellite DNA, prevailing few variants. On the contrary, the contraction events that involve tandems of homogeneous monomers contribute—by default—minor variants to become “evident”, which otherwise were undetectable. On the other hand, all the RPCS sequence variants are totally or partially shared by all the studied populations. As a whole, these results are comprehensible if these RPCS variants preexisted in the common ancestor of this Ctenomys group.Fil: Caraballo, Diego Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Belluscio, Pablo Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Rossi, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin