SummaryThe MerR family is a group of bacterial transcriptional regulators that respond to different environmental stimuli, such as heavy metals, oxidative stress or antibiotics. Here we characterize a new member of this family that is highly selective for Au ions. We show that this Salmonella regulator, named GolS, directly controls the expression of at least two transcriptional units specifically required for Au resistance. By chromosomal mutagenesis, we demonstrated that Au-selectivity is accomplished by a metal-binding motif in GolS. Among the monovalent metal-ion sensing MerR regulators GolS clusters in a branch distant from enterobacterial CueR orthologues. We propose that GolS and its homologues evolved to cope with toxic concentration of Au ion, allowing microorganisms to withstand contaminated environments.

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The mismatch repair (MMR) system maintains genome integrity by correcting replication-associated errors and inhibiting recombination between divergent DNA sequences. The basic features of the pathway have been highly conserved throughout evolution, although the nature and number of the proteins involved in this DNA repair system vary among organisms. Plants have an extra mismatch recognition protein, MutSγ, which is a heterodimer: MSH2-MSH7. To further understand the role of MSH7 in vivo, we present data from this protein in Arabidopsis thaliana. First, we generated transgenic plants that express β-glucuronidase (GUS) under the control of the MSH7 promoter. Histochemical staining of the transgenic plants indicated that MSH7 is preferentially expressed in proliferating tissues. Then, we identified msh7 T-DNA insertion mutants. Plants deficient in MSH7 show increased levels of UV-B-induced cyclobutane pyrimidine dimers relative to wild-type (WT) plants. Consistent with the patterns of MSH7 expression, we next analysed the role of the protein during somatic and meiotic recombination. The frequency of somatic recombination between homologous or homeologous repeats (divergence level of 1.6%) was monitored using a previously described GUS recombination reporter assay. Disruption of MSH7 has no effect on the rates of somatic homologous or homeologous recombination under control conditions or after UV-B exposure. However, the rate of meiotic recombination between two genetically linked seed-specific fluorescent markers was 97% higher in msh7 than in WT plants. Taken together, these results suggest that MSH7 is involved in UV-B-induced DNA damage recognition and in controlling meiotic recombination.

SUMMARYCurrently, the storage of fish spermatozoa through cryopreservation is widely used. Although it is of common knowledge that the process of freezing / thawing generates DNA fragmentation of spermatozoa, the consequences of this process on embryonary development are unknown. There is great interest in developing methods for in vitro fertilization for the species Prochilodus lineatus using cryopreserved semen. In this paper we study the embryonic development of this fish, to lay the groundwork for the observation of abnormalities in the development of embryos derived from criopreserved spermatozoa. The eggs produced by this species are translucent and have a large perivitelline space, are of the telolecitic type, and presented meroblastic division during the early stages of development. Although the embrionary development of Prochilodus lineatus (Sábalo) took place in less time compared with Danio rerio (Zebrafish) the embryo goes through very similar stages. During the study it was possible to observe the periods of zygote, cleavage, blastula, gastrula, segmentation and hatching of the larvae of Sábalo. The different stages of development were successfully recorded, especially those after the transition of blastula media (TBM), when the embryo begins to transcribe its own genome. Palabras clave: Prochilodus lineatus, peces, embriones.Key words: Prochilodus lineatus, fish, embryos. INTRODUCCIÓN(Ninhaus-Silveira y col 2006). En Brasil existen pocos estudios acerca del desarrollo embrionario del sábalo El sábalo (Prochilodus lineatus) es un pez de la familia (Prochilodus lineatus). La descripción del desarrollo Prochilodontidae, iliófago, distribuido por todo el sudeste embrionario puede tener innumerables ventajas, como favode Brasil, que migra para desovar durante el período de recer el reconocimiento de los embriones en sus ambientes reproducción (Fowler 1950). Esta es una especie encontrada naturales, permitiendo una mejor evaluación del sitio de por toda la cuenca del Río Paraná-Paraguay y Paraíba y desove de los peces y estudios relativos al crecimiento de tiene gran importancia económica y ecológica entre las la especie en su ambiente natural; también es importante especies nativas de tamaño medio y grande. para la detección de las alteraciones relacionadas con los Existen diversas especies de peces en Brasil que han factores ambientales en las incubadoras, que pueden llevar sido poco exploradas y para muchas de éstas se desconoce a una malformación de las larvas y consecuentemente una el potencial zootécnico. Para obtener éxito en la producmenor productividad (Alves y Moura 1992). Las invesción acuícola se vuelve necesario el conocimiento de las tigaciones relativas al desarrollo embrionario y larval en características morfofisiológicas y conductuales de las peces en cautiverio han sido realizadas principalmente en especies en estudio, siendo por este motivo de vital imespecies de interés comercial (Luz y col 2001, Romagosa portancia el estudio de los primeros días de vida (Pezzato y col 2001). 1997). ...

In order to improve the understanding of pejerrey Odontesthes bonariensis, growth hormone (Gh)-insulin-like growth factor-1(Igf1) axis, O. bonariensis growth hormone receptor type 1 (ghr1) and type 2 (ghr2) mRNA sequences were obtained. Both transcripts were ubiquitously expressed except in kidney, encephalon and anterior intestine. Alternative transcripts of both receptors were found in muscle. Interestingly, two different ghr2 transcripts with alternative polyadenylation (APA) sites located in the long 3' untranslated region (UTR-APA) were also found in liver. Hepatic ghr1, ghr2 and insulin-like growth factor type 1 (igf1) transcript levels were examined under two different metabolic conditions. In the first experimental condition, fish were fasted for 2 weeks and then re-fed for another 2 weeks. Despite igf1 mRNA relative expression did not show significant differences under the experimental period of time examined, both ghr transcripts decreased their expression levels after the fasting period and returned to their control levels after re-feeding. In the second treatment, recombinant O. bonariensis growth hormone (r-pjGh) was orally administered once a week. After 4 weeks of treatment, liver igf1, ghr1 and ghr2 mRNA relative expression increased (13, 4·5 and 2·1 fold, P < 0·05) compared to control values. These results add novel information to the growth hormone-insulin-like growth factor system in teleosts.

Using biotechnology to increase the growth rates of fish is likely to reduce production costs per unit of food. Among vertebrates, fish appear to occupy a unique position, when growth patterns are considered. With few exceptions, fish species tend to grow indeterminately, implying that size is never fixed. Both hyperplasia and hypertrophy contribute to post-larval muscle growth in fish. Growth hormone (GH) - Insulin-like Growth Factor I (IGF-I) is the most important growth axis in fish. Our experimental model, the pejerrey, Odontesthes bonariensis (Ateriniformes) is a South American inland water fish considered to be a promising species for intensive aquaculture. However, one major drawback to achieve this goal is its slow growth in captivity. In order to understand how growth is regulated in this species, our first objective was to characterized pejerrey GH- IGF-I axis. We first cloned and characterized pejerrey (pj) GH, IGF-I and the growth hormone receptors (GHRs) I and II. In addition to providing valuable data for evolutionary comparison of GH, investigation of GH action in teleosts is particularly important because of its potential application in aquaculture. GH can not only promote the somatic growth in fish but also lower dietary protein requirements. A prerequisite for providing sufficient amounts of GH for basic research and aquaculture application is a large-scale production of GH. For that purpose, recombinant pjGH was expressed in a bacterial system. Protocols for solubilization and proper folding were achieved. Activity of recombinant pjGH was assessed in fish by measuring the liver IGF-I response to different doses of GH. IGF-I transcript was measured in the liver after pjGHr in vivo stimulation by means of quantitative real-time PCR assays. A dose-dependent response of IGF-I mRNA was observed after pjGHr administration, and reached a 6 fold IGF-I maximum increase over control group when 2.5 µg pjGH /g-body weight were injected. Temporal analysis of hepatic IGF-I mRNA levels showed that administration of a single dose of pjGHr into juvenile pejerrey resulted in a significant increase (P<0.02) 9 hours post injection. These results demonstrates that recombinant pjGH could promote a dramatic response in liver, increasing the IGF-I mRNA level. We also study the effect of GH on muscle growth after oral administration. A significant association between GH doses and mean fiber area (MFA) was observed even with a caloric restrictive diet. MFA increased 3.7 µm² per each unit of GH supplied indicating that GH promoted white muscle hypertrophy. These preliminary data indicates that GH could be absorbed by the intestine in an active form and promote somatic growth

Glycosidases are present both in sperm and eggs in vertebrates and have been associated with different fertilization steps as gamete binding, egg coat penetration, and polyspermy prevention. In this manuscript, we have analyzed the activity of different glycosidases of Xenopus laevis eggs. The main activity corresponded to N-acetyl-β-D-glucosaminidase (Hex), which was reported to participate both in gamete binding and polyspermy prevention among phylogenetically distant animals. We have raised homologous antibodies against a recombinant N-terminal fragment of a X. laevis Hex, and characterized egg's Hex both by Western blot and immunohistochemical assays. Noteworthy, Hex was mainly localized to the cortex of animal hemisphere of full-grown oocytes and oviposited eggs, and remained unaltered after fertilization. Hex is constituted by different pair arrangements of two subunits (α and β), giving rise to three possible Hex isoforms: A (αβ), B (ββ), and S (αα). However, no information was available regarding molecular identity of Hex in amphibians. We present for the first time the primary sequences of two isoforms of X. laevis Hex. Interestingly, our results suggest that α- and β-like subunits that constitute Hex isoforms could be synthesized from a same gene in Xenopus, by alternative exon use. This finding denotes an evolutionary divergence with mammals, where α and β Hex subunits are synthesized from different genes on different chromosomes.