RNA-binding protein Maca is crucial for gigantic male fertility factor gene expression, spermatogenesis, and male fertility, in Drosophila

During spermatogenesis, the process in which sperm for fertilization are produced from germline cells, gene expression is spatiotemporally highly regulated. In Drosophila, successful expression of extremely large male fertility factor genes on Y-chromosome spanning some megabases due to their gigantic intron sizes is crucial for spermatogenesis. Expression of such extremely large genes must be challenging, but the molecular mechanism that allows it remains unknown. Here we report that a novel RNA-binding protein Maca, which contains two RNA-recognition motifs, is crucial for this process. maca null mutant male flies exhibited a failure in the spermatid individualization process during spermatogenesis, lacked mature sperm, and were completely sterile, while maca mutant female flies were fully fertile. Proteomics and transcriptome analyses revealed that both protein and mRNA abundance of the gigantic male fertility factor genes kl-2, kl-3, and kl-5 (kl genes) are significantly decreased, where the decreases of kl-2 are particularly dramatic, in maca mutant testes. Splicing of the kl-3 transcripts was also dysregulated in maca mutant testes. All these physiological and molecular phenotypes were rescued by a maca transgene in the maca mutant background. Furthermore, we found that in the control genetic background, Maca is exclusively expressed in spermatocytes in testes and enriched at Y-loop A/C in the nucleus, where the kl-5 primary transcripts are localized. Our data suggest that Maca increases transcription processivity, promotes successful splicing of gigantic introns, and/or protects transcripts from premature degradation, of the kl genes. Our study identified a novel RNA-binding protein Maca that is crucial for successful expression of the gigantic male fertility factor genes, spermatogenesis, and male fertility.

ARTISTIC EXPRESSION OF CARICATURES OF PEOPLE'S ARTIST HUSEYNGULU ALIYEV

In Huseyngulu Aliyev's works, we always see the successful expression of harsh color transitions, tonal spots, light effects, instant glare in the formation of dynamism, as well as the artist's unique creative position with different compositional structures in the more exciting presentation of these effects. In his work "Molla Nasreddin", which is currently preserved in a private collection in Norway, the artist expressed the deep expression of dynamism in the composition, both in the intensity of movements, sudden jumps, hymns of speed, and the contrasting transition of colors from light to dark or vice versa. It is interesting to recreate the artistic solution of the instantaneous plot in which the fruit of the old man, sitting on a fast-running donkey, spreads in the air and begins to scatter in the air. The clear language of caricatures created by Huseyngulu Aliyev, which combines both matte and bright tones of colors such as green, yellow and brown, is very thought-provoking due to their mobility and dynamism. Created in 1993, the interesting language of expression of the works, which is a vivid example of the idea mentioned in the cartoons with ink and pen, attracts attention. In "Unsuccesfull" the intention of a thief in a black mask to plunder the country, but before that the looting of these places ended in the failure of his plan. The astonishment of the robber looking at the empty baskets and glasses scattered on the ground makes it possible to imagine the facial features he covers. The clear language of caricatures created by Huseyngulu Aliyev is also very thought-provoking due to their mobility and dynamism. Created in 1993, the interesting language of expression of the works, which is a vivid example of the idea mentioned in the cartoons with ink and pen, attracts attention. In "Unsuccesfull" the intention of a thief in a black mask to plunder the country, but before that the looting of these places ended in the failure of his plan.The astonishment of the robber looking at the empty baskets and glasses scattered on the ground makes it possible to imagine the facial features he covers. Among the graphic examples created in the mentioned period, the unique composition of the work "Roads" attracts the attention of the audience with its interesting composition, which has a great meaning. The artist sang the song of a long way with the movement of crowded people in the same direction. The march of people who join the movement on an empty background expresses their common thinking and will. Here, the artist has successfully implemented mass thinking, not where and why the roads go. It is well-known that art, which came to art by chance, but managed to introduce itself in any way, has gained popularity in our time. The artist's very interesting and humorous composition in his caricature "Patriot of art" created about 20 years ago is dedicated to this type of "artists". The stage hymn of the performance of a long-eared man holding his tambourine in his hands in an artistic form clearly expresses the artist's purpose. The solution of the symbolic meaning given to some scenes of life by the artist using folk sayings in the unique compositional structure gives the basis to evaluate the artist's creativity by presenting it in a different form in each work. For example, in his 2001 work, “They Make the Old Moon a Star”, he praised an astrologer sitting on his back with his head cut off and turning the old moon into a star in the sky. The artist was referring to people who were ungrateful and apostate (Figure 2.91). In Huseyngulu Aliyev's caricatures, artistic exaggerations, different assessments of events or directing them in accordance with the content of the work are of interest as creative extraordinary creative discoveries. For example, the artist, who turned a fragment from the world-famous sculptor Auguste Rodin's "The Thinker" into the content of the painting, watched the two snakes collide, fight, and try to poison each other, thinking deeply about world events. Here, the sharpness of the artistic generalization in such a difficult scene as the destruction of the same sex makes the viewer think (Figure 2.96).

Pinpointing Synechococcus Rubisco Large Subunit Sections Involved in Heterologous Holoenzyme Formation in Escherichia Coli

Background: Heterologous holoenzyme formation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) have been a challenge due to limited understanding of its biogenesis. Unlike bacterial Rubiscos, eukaryotic Rubiscos are incompatible with the Escherichia coli chaperone system to fold and assemble into the functional hexadecameric conformation (L8S8), which comprise eight large subunits (RbcL) and eight small subunits (RbcS). Our previous study reported three sections (residues 248-297, 348-397 and 398-447) within the RbcL of Synechococcus elongatus PCC6301 may be important for formation of L8S8 in E. coli. Present study further examined these three sections separately, by dividing them into six sections of 25 residues (i.e. residues 248-272, 273-297, 348-372, 373-397, 398-422 and 423-447). Methods and Results: Six chimeric Rubiscos with each section within the RbcL from Synechococcus replaced by their respective counterpart sequence from Chlamydomonas reinhardtii were constructed and checked for their effect on holoenzyme formation in E. coli. Present study shows that Section 1 (residues 248-272; section of Synechococcus RbcL replaced by corresponding Chlamydomonas sequence), Section 2 (residues 273-297), Section 3 (residues 348-372) and Section 6 (residues 423-447) chimeras failed to fold and/or assemble despite successful expression of both RbcL and RbcS. Only Section 4 (residues 373-397) and 5 (residues 398-422) chimeras could form L8S8 in E. coli. Conclusion: As GroEL chaperonin mediates folding of bacterial RbcL in E. coli, residues 248-297, 348-372 and 423-447 of Synechococcus RbcL may be important for interacting with the GroEL chaperonin for successful holoenzyme formation in E. coli.

The Wild Worm Codon Adapter: a web tool for automated codon adaptation of transgenes for expression in non-Caenorhabditis nematodes

ABSTRACTAdvances in genomics techniques are expanding the range of nematode species that are amenable to transgenesis. Due to divergent codon usage biases across species, codon optimization is often a critical step for the successful expression of exogenous transgenes in nematodes. Platforms for generating DNA sequences codon optimized for the free-living model nematode Caenorhabditis elegans are broadly available. However, until now such tools did not exist for non-Caenorhabditis nematodes. We therefore developed the Wild Worm Codon Adapter, a tool for rapid transgene codon optimization for expression in non-Caenorhabditis nematodes. The app includes built-in optimization for parasitic nematodes in the Strongyloides, Nippostrongylus and Brugia genera as well as the predatory nematode Pristionchus pacificus. The app also supports custom optimization for any species using user-provided optimization rules. In addition, the app supports automated insertion of synthetic or native introns, as well as the analysis of codon bias in transgene and native sequences. Here, we describe this web-based tool and demonstrate how it may be used to analyze genome-wide codon bias in Strongyloides species.

Genomic deletions in Rhodococcus based on transformation of linear heterologous DNA

Several genome engineering methods have been developed for Rhodococcus . However, they suffer from limitations such as extensive cloning, multiple steps, successful expression of heterologous genes via plasmid etc. Here, we report a rapid method for performing genomic deletions/disruptions in Rhodococcus spp. using heterologous linear DNA. The method is cost effective and less labour intensive. The applicability of the method was demonstrated by successful disruption of rodA and orphan parA. None of the disrupted genes were found to be essential for the viability of the cell. Disruption of orphan parA and rodA resulted in elongated cells and short rods, respectively. This is the first report demonstrating disruption of rodA and orphan parA genes by electroporation of heterologous linear DNA in Rhodococcus spp.

Efficient biomass saccharification using a novel cellobiohydrolase from Clostridium clariflavum for utilization in biofuel industry

Successful expression of a novel cellobiohydrolase enzyme from Clostridium clariflavum with efficient saccharification potential of plant biomass for the biofuel industry.

Making, Cloning and Expression of Human Insulin Genes in Bacteria: The Path to Humulin@

In the mid to late 1970s recombinant DNA methods for cloning and expressing genes in E. coli were under intense development. An important question had become: Can humans design and chemically synthesize novel genes that function in bacteria? This question was answered in 1978 and 1979 with the successful expression in E. coli of two mammalian hormones, first somatostatin and then human insulin. The successful production of human insulin in bacteria provided, for the first time, a practical, scalable source of human insulin and resulted in the approval, in 1982, of human insulin for the treatment of diabetics. In this short review, I give my personal view of how the making, cloning and expressing of human insulin genes was accomplished by a team of scientists led by Keiichi Itakura, Herbert W. Boyer and myself.

3-D Nucleus Architecture in Oat × Maize Addition Lines

The nucleus architecture of hybrid crop plants is not a well-researched topic, yet it can have important implications for their genetic stability and usefulness in the successful expression of agronomically desired traits. In this work we studied the spatial distribution of introgressed maize chromatin in oat × maize addition lines with the number of added maize chromosomes varying from one to four. The number of chromosome additions was confirmed by genomic in situ hybridization (GISH). Maize chromosome-specific simple sequence repeat (SSR) markers were used to identify the added chromosomes. GISH on 3-D root and leaf nuclei was performed to assess the number, volume, and position of the maize-chromatin occupied regions. We revealed that the maize chromosome territory (CT) associations of varying degree prevailed in the double disomic lines, while CT separation was the most common distribution pattern in the double monosomic line. In all analyzed lines, the regions occupied by maize CTs were located preferentially at the nuclear periphery. A comparison between the tissues showed that the maize CTs in the leaf nuclei are positioned closer to the center of the nucleus than in the root nuclei. These findings shed more light on the processes that shape the nucleus architecture in hybrids.

The Memorial ACTe

This essay examines the nature, scope and consequences of the seism of memory since its eruption in 2000 in Guadeloupe, French West Indies. In particular, it questions how the context of a multifaceted appetite for collective remembrance took the form of competing strategies for memorialization in the space. As such, it focuses on the heritage of pain, resistance and pride at the local, national and regional levels. I draw on Shalini Puri’s analysis of the repressed memory of the 1983 Grenada revolution in Operation Urgent Memory to identify in the landscape of Guadeloupe submerged, residual and eruptive ‘platforms of memory’ (Puri, 2012). In the specific case of Guadeloupe, the collective efforts of the Guadeloupean people for re-appropriating their non-French and non-European heritage on the island have turned into competitive post-traumatic approaches of the history of transatlantic slave trade. This essay eventually analyses the case of the Mémorial ACTe (MACTe) – Museum of Contemporary Caribbean Art and Memorial for the History of the Slave Trade – as constituting the most successful expression of what I define as cultural marronage, in the ambivalent postcolonial environment of the French Overseas Regions.