scholarly journals Effects of Osiris9a on Silk Properties in Bombyx mori Determined by Transgenic Overexpression

2020 ◽  
Vol 21 (5) ◽  
pp. 1888
Author(s):  
Tingcai Cheng ◽  
Xia Zhang ◽  
Zhangchuan Peng ◽  
Yinfeng Fan ◽  
Lin Zhang ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Biological Function ◽  
Evolutionary Conservation ◽  
Silk Gland ◽  
Fiber Formation ◽  
Silk Fiber ◽  
Specific Gene ◽  
Silk Proteins ◽  
Insight Into ◽  
Posterior Silk Gland

Osiris is an insect-specific gene family with multiple biological roles in development, phenotypic polymorphism, and protection. In the silkworm, we have previously identified twenty-five Osiris genes with high evolutionary conservation and remarkable synteny among several insects. Bombxy mori Osiris9a (BmOsi9a) is expressed only in the silk gland, particularly in the middle silk gland (MSG). However, the biological function of BmOsi9a is still unknown. In this study, we overexpressed BmOsi9a in the silk gland by germline transgene expression. BmOsi9a was overexpressed not only in the MSG but also in the posterior silk gland (PSG). Interestingly, BmOsi9a could be secreted into the lumen in the MSG but not in the PSG. In the silk fiber, overexpressed BmOsi9a interacted with Sericin1 in the MSG, as confirmed by a co-immunoprecipitation assay. The overexpression of BmOsi9a altered the secondary structure and crystallinity of the silk fiber, thereby changing the mechanical properties. These results provide insight into the mechanisms underlying silk proteins secretion and silk fiber formation.

Phosphoproteomic analysis of the posterior silk gland of Bombyx mori provides novel insight into phosphorylation regulating the silk production

2016 ◽  
Vol 148 ◽  
pp. 194-201 ◽  
Author(s):  
Jia Song ◽  
Jiaqian Che ◽  
Zhengying You ◽  
Lupeng Ye ◽  
Jisheng Li ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Silk Gland ◽  
Silk Production ◽  
Insight Into ◽  
Posterior Silk Gland

scholarly journals Deep Insight into the Transcriptome of the Single Silk Gland of Bombyx mori

2019 ◽  
Vol 20 (10) ◽  
pp. 2491 ◽  
Author(s):  
Run Shi ◽  
Sanyuan Ma ◽  
Ting He ◽  
Jian Peng ◽  
Tong Zhang ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Fatty Acid ◽  
Fatty Acid Biosynthesis ◽  
Silk Gland ◽  
Fiber Formation ◽  
Silk Fiber ◽  
Silk Protein ◽  
High Concentration ◽  
And Function ◽  
Source Use

The silk gland synthesizes and secretes a large amount of protein and stores liquid silk protein at an extremely high concentration. Interestingly, silk proteins and serine protease inhibitors are orderly arranged in the silk gland lumen and cocoon shells. Silk fiber formation and the spinning mechanism have not been fully elucidated. Therefore, we conducted a comparative transcriptome analysis of seven segments of the single silk gland to characterize internal changes in the silk gland during the 5th instar of mature larvae. In total, 3121 differentially expressed genes were identified in the seven segments. Genes highly expressed in the middle silk gland (MSG) were mainly involved in unsaturated fatty acid biosynthesis, fatty acid metabolism, apoptosis—fly, and lysosome pathways, whereas genes highly expressed in the posterior silk gland (PSG) were mainly involved in ribosome, proteasome, citrate cycle, and glycolysis/gluconeogenesis pathways. Thus, the MSG and PSG differ greatly in energy source use and function. Further, 773 gradually upregulated genes (from PSG to MSG) were involved in energy metabolism, silk protein synthesis, and secretion, suggesting that these genes play an important role in silk fiber formation. Our findings provide insights into the mechanism of silk protein synthesis and transport and silk fiber formation.

scholarly journals Enhanced Myc Expression in Silkworm Silk Gland Promotes DNA Replication and Silk Production

Insects ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 361
Author(s):  
Wenliang Qian ◽  
Yan Yang ◽  
Zheng Li ◽  
Yuting Wu ◽  
Xuechuan He ◽  
...  
Keyword(s):  
Dna Replication ◽  
Cell Growth ◽  
Silk Gland ◽  
Biological Functions ◽  
Gland Cells ◽  
Silk Production ◽  
Myc Gene ◽  
Cell Growth And Proliferation ◽  
Silkworm Silk ◽  
Posterior Silk Gland

Silkworm is an economically important insect that synthetizes silk proteins for silk production in silk gland, and silk gland cells undergo endoreplication during larval period. Transcription factor Myc is essential for cell growth and proliferation. Although silkworm Myc gene has been identified previously, its biological functions in silkworm silk gland are still largely unknown. In this study, we examined whether enhanced Myc expression in silk gland could facilitate cell growth and silk production. Based on a transgenic approach, Myc was driven by the promoter of the fibroin heavy chain (FibH) gene to be successfully overexpressed in posterior silk gland. Enhanced Myc expression in the PSG elevated FibH expression by about 20% compared to the control, and also increased the weight and shell rate of the cocoon shell. Further investigation confirmed that Myc overexpression increased nucleus size and DNA content of the PSG cells by promoting the transcription of the genes involved in DNA replication. Therefore, we conclude that enhanced Myc expression promotes DNA replication and silk protein expression in endoreplicating silk gland cells, which subsequently raises silk yield.

scholarly journals Simulation of Folding Kinetics for Aligned RNAs

Genes ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 347
Author(s):  
Jiabin Huang ◽  
Björn Voß
Keyword(s):  
Evolutionary Conservation ◽  
Folding Kinetics ◽  
New Approach ◽  
Computational Approaches ◽  
Reasonable Time ◽  
Valuable Method ◽  
Kinetics Of ◽  
Insight Into ◽  
Folding Space ◽  
Standing Problem

Studying the folding kinetics of an RNA can provide insight into its function and is thus a valuable method for RNA analyses. Computational approaches to the simulation of folding kinetics suffer from the exponentially large folding space that needs to be evaluated. Here, we present a new approach that combines structure abstraction with evolutionary conservation to restrict the analysis to common parts of folding spaces of related RNAs. The resulting algorithm can recapitulate the folding kinetics known for single RNAs and is able to analyse even long RNAs in reasonable time. Our program RNAliHiKinetics is the first algorithm for the simulation of consensus folding kinetics and addresses a long-standing problem in a new and unique way.

scholarly journals STUDIES ON THE POSTERIOR SILK GLAND OF THE SILKWORM BOMBIX MORI

1970 ◽  
Vol 46 (1) ◽  
pp. 1-16 ◽  
Author(s):  
Yutaka Tashiro ◽  
Eiichi Otsuki
Keyword(s):  
Guanidine Hydrochloride ◽  
Lithium Bromide ◽  
Posterior Part ◽  
Silk Gland ◽  
Sedimentation Equilibrium ◽  
Partial Specific Volume ◽  
Anterior Portion ◽  
Equilibrium Analyses ◽  
Fibroin Solution ◽  
Posterior Silk Gland

Ultracentrifugal analyses of the native silk proteins extracted from the various parts of the middle silk gland of the mature silkworm have revealed that there exist four components with S°20,w values of 10S, 9–10S, 9S, and 4S in the extract. It is suggested that the fastest 10S component is the native fibroin synthesized in the posterior silk gland and transferred to the middle silk gland to be stored there, while the slower three components probably correspond to inner, middle, and outer sericins which were synthesized in the posterior, middle, and anterior portion of the middle silk gland, respectively. Native fibroin solution was prepared from the most posterior part of the middle silk gland. Ultracentrifugal analyses have shown that the solution contains considerable amounts of aggregates in addition to the main 10S component. Treatment with lithium bromide (LiBr), urea, or guanidine hydrochloride solution up to 6 M all have failed to dissociate the 10S component. From the sedimentation equilibrium analyses and partial specific volume of 0.716, the molecular weight of the 10S component of the native fibroin solution was found to be between 3.2 – 4.2 x 105, with a tendency to lie fairly close to 3.7 x 105.

Participation of the upstream region of the fibroin gene in the formation of transcription complex in vitro

1986 ◽  
Vol 6 (11) ◽  
pp. 3928-3933
Author(s):  
M Tsuda ◽  
S Hirose ◽  
Y Suzuki
Keyword(s):  
Complex Formation ◽  
Specific Interaction ◽  
Inhibitory Effect ◽  
Silk Gland ◽  
Upstream Region ◽  
Fibroin Gene ◽  
Transcription Complexes ◽  
Posterior Silk Gland

The addition of exogenous histones has an inhibitory effect on fibroin gene transcription in posterior silk gland extracts. The histones probably disturb a process in complex formation, because when transcription complexes were constructed by preincubation of the templates with the extracts, the inhibitory effect of histones was greatly reduced. Transcription of a fibroin gene construct, pFb5' delta-238, having the upstream region beyond the TATA box was relatively less inhibited than that of pFb5' delta-44 lacking the upstream region. This tendency toward differential inhibition was observed in the silk gland extracts but not in a HeLa cell extract and persisted even after complex formation in the silk gland extracts, suggesting a specific interaction of the upstream region with some factors in the extracts. The complexes formed on pFb5' delta-44 are probably more susceptible to the inhibitory effect of histones. On the basis of these results we propose a participation of the upstream region of the fibroin gene in the formation of stable transcription complexes at the promoter through an interaction with specific factors in the silk gland. Since the transcription-enhancing effect via the upstream region is augmented at a high histone/DNA ratio, it may mimic the in vivo situation in which the fibroin gene can be transcribed in the posterior silk gland even in the presence of excess suppressive materials.

scholarly journals Transgenic Epigenetics: Using Transgenic Organisms to Examine Epigenetic Phenomena

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
Lori A. McEachern
Keyword(s):  
Molecular Mechanisms ◽  
Model Organism ◽  
Evolutionary Conservation ◽  
Model Organisms ◽  
Valuable Insight ◽  
Epigenetic Control ◽  
Transgenic Organisms ◽  
Epigenetic Analysis ◽  
Insight Into ◽  
Epigenetic Processes

Non-model organisms are generally more difficult and/or time consuming to work with than model organisms. In addition, epigenetic analysis of model organisms is facilitated by well-established protocols, and commercially-available reagents and kits that may not be available for, or previously tested on, non-model organisms. Given the evolutionary conservation and widespread nature of many epigenetic mechanisms, a powerful method to analyze epigenetic phenomena from non-model organisms would be to use transgenic model organisms containing an epigenetic region of interest from the non-model. Interestingly, while transgenic Drosophila and mice have provided significant insight into the molecular mechanisms and evolutionary conservation of the epigenetic processes that target epigenetic control regions in other model organisms, this method has so far been under-exploited for non-model organism epigenetic analysis. This paper details several experiments that have examined the epigenetic processes of genomic imprinting and paramutation, by transferring an epigenetic control region from one model organism to another. These cross-species experiments demonstrate that valuable insight into both the molecular mechanisms and evolutionary conservation of epigenetic processes may be obtained via transgenic experiments, which can then be used to guide further investigations and experiments in the species of interest.

scholarly journals Photoperiodic modulation of circadian rhythms in the silk gland protein profiles of Bombyx mori and its influence on the silk productivity and quality

2010 ◽  
Vol 2 (1) ◽  
pp. 48-56 ◽  
Author(s):  
B. Sailaja ◽  
S. Sivaprasad
Keyword(s):  
Circadian Rhythms ◽  
Bombyx Mori ◽  
Continuous Light ◽  
Silk Gland ◽  
Protein Profiles ◽  
Dark Cycle ◽  
Response Curves ◽  
Silk Proteins ◽  
Free Running ◽  
Clock Shift

Circadian rhythms in the silk gland protein profiles of Bombyx mori were analyzed under 12 h light and 12 h dark cycle (LD), continuous light (LL) and continuous dark (DD) conditions. The phase response curves of protein rhythms indicate the prevalence of a series of silk cycles, each comprising three phases; transcription, translation and consolidation of silk proteins. In the 24h- protein rhythm, the silk cycle repeats every 3h, 42 m under LD, 2h, 36m under LL and 3h under DD. The light and dark conditions advanced the rhythm of each silk cycle by 48m and 24m respectively. As a result the silk gland completes 7 rounds of protein synthesis under LD, 9 rounds under LL and 8 rounds under DD during the 24h-free running time of the rhythm. The light-induced clock-shift in the protein rhythm caused significant gains in economic parameters of sericulture with positive signals for enhancing silk productivity and quality.

scholarly journals FUS-dependent loading of SUV39H1 to OCT4 pseudogene-lncRNA programs a silencing complex with OCT4 promoter specificity

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Michele Scarola ◽  
Elisa Comisso ◽  
Massimo Rosso ◽  
Giannino Del Sal ◽  
Claudio Schneider ◽  
...  
Keyword(s):  
Rna Binding ◽  
Biological Function ◽  
Rna Binding Protein ◽  
Evolutionary Conservation ◽  
Epigenetic Silencing ◽  
Site Specificity ◽  
Sequence Element ◽  
Regulatory Circuits ◽  
Promoter Specificity ◽  
Dependent Mechanism

Abstract The resurrection of pseudogenes during evolution produced lncRNAs with new biological function. Here we show that pseudogene-evolution created an Oct4 pseudogene lncRNA that is able to direct epigenetic silencing of the parental Oct4 gene via a 2-step, lncRNA dependent mechanism. The murine Oct4 pseudogene 4 (mOct4P4) lncRNA recruits the RNA binding protein FUS to allow the binding of the SUV39H1 HMTase to a defined mOct4P4 lncRNA sequence element. The mOct4P4-FUS-SUV39H1 silencing complex holds target site specificity for the parental Oct4 promoter and interference with individual components results in loss of Oct4 silencing. SUV39H1 and FUS do not bind parental Oct4 mRNA, confirming the acquisition of a new biological function by the mOct4P4 lncRNA. Importantly, all features of mOct4P4 function are recapitulated by the human hOCT4P3 pseudogene lncRNA, indicating evolutionary conservation. Our data highlight the biological relevance of rapidly evolving lncRNAs that infiltrate into central epigenetic regulatory circuits in vertebrate cells.

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