The Evolutionary Conserved Transmembrane BAX Inhibitor Motif (TMBIM) Containing Protein Family Members 5 and 6 Are Essential for the Development and Survival of Drosophila melanogaster

The mammalian Transmembrane BAX Inhibitor Motif (TMBIM) protein family consists of six evolutionarily conserved hydrophobic proteins that affect programmed cell death and the regulation of intracellular calcium levels. The bacterial ortholog BsYetJ is a pH-dependent calcium channel. We here identified seven TMBIM family members in Drosophila melanogaster and describe their expression levels in diverse tissues and developmental stages. A phylogenetic analysis revealed that CG30379 represents the ortholog of human TMBIM4 although these two proteins are much less related than TMBIM5 (CG2076 and CG1287/Mics1) and TMBIM6 (CG7188/Bi-1) to their respective orthologs. For TMBIM1-3 the assignment is more dubious because the fly and the human proteins cluster together. We conducted a functional analysis based on expression levels and the availability of RNAi lines. This revealed that the ubiquitous knockdown of CG3798/Nmda1 and CG3814/Lfg had no effect on development while knockdown of CG2076/dTmbim5 resulted in death at the pupa stage and knockdown of CG7188/dTmbim6 in death at the embryonic stage. Ubiquitous knockdown of the second TMBIM5 paralog CG1287/Mics1 ensued in male sterility. Knockdown of dTmbim5 and 6 in muscle and neural tissue also greatly reduced lifespan through different mechanisms. Knockdown of the mitochondrial family member dTmbim5 resulted in reduced ATP production and a pro-apoptotic expression profile while knockdown of the ER protein dTmbim6 increased the ER calcium levels similar to findings in mammalian cells. Our data demonstrate that dTmbim5 and 6 are essential for fly development and survival but affect cell survival through different mechanisms.

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The Evolutionary Conserved Transmembrane BAX Inhibitor Motif (TMBIM) Containing Protein Family is Essential for the Development and Survival of Drosophila Melanogaster

10.21203/rs.3.rs-113374/v1 ◽

2020 ◽

Author(s):

Li Zhang ◽

Sebastian Buhr ◽

Vibha Prasad ◽

Aaron Voigt ◽

Axel Methner

Keyword(s):

Drosophila Melanogaster ◽

Genetic Interaction ◽

Family Members ◽

Protein Family ◽

Tissue Specific ◽

Similar Expression Pattern ◽

Evolutionarily Conserved ◽

Extended Lifespan ◽

Ph Dependent ◽

Er Calcium

Abstract The Transmembrane BAX Inhibitor Motif (TMBIM) protein family consists of six evolutionarily conserved hydrophobic proteins that affect programmed cell death and the regulation of intracellular calcium levels. The bacterial orthologue BsYetJ is a pH-dependent calcium channel. We here identified six TMBIM family members in Drosophila melanogaster and studied the effect of their RNAi-mediated knockdown using ubiquitous and tissue-specific drivers. Mammalian TMBIM6 and TMBIM5 have obvious orthologs while this is more dubious for the other family members. Ubiquitous knockdown of family members dmTMBIM1,4,5, and 6 caused failed eclosing and tissue-specific knockdown resulted in a dramatically decreased lifespan. On the contrary, knockdown of dmTMBIM3, surprisingly, extended lifespan. Only knockdown of dmTMBIM6 increased the ER calcium levels of Pdf neurons. Neural knockdown of dmTMBIM2,3, and 4 increased ER stress, as indicated by increased Xbp1 splicing. Interestingly, TMBIM1 and TMBIM6 have a very similar expression pattern and their knockdown phenocopied each other. Also, knockdown of TMBIM1 resulted in upregulation of TMBIM6 and vice versa further suggesting a genetic interaction between these two genes. Our data demonstrate that most TMBIM proteins are essential for fly development and survival but, despite their shared protein structure, affect cell survival through different mechanisms.

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Apoptosis and development

Essays in Biochemistry ◽

10.1042/bse0390011 ◽

2003 ◽

Vol 39 ◽

pp. 11-24 ◽

Cited By ~ 6

Author(s):

Justin V McCarthy

Keyword(s):

Drosophila Melanogaster ◽

Mammalian Cells ◽

Cell Number ◽

Model Organisms ◽

Biological Processes ◽

Nematode Caenorhabditis Elegans ◽

Apoptotic Pathway ◽

Genetic Pathways ◽

Evolutionarily Conserved ◽

Multicellular Organisms

Apoptosis is an evolutionarily conserved process used by multicellular organisms to developmentally regulate cell number or to eliminate cells that are potentially detrimental to the organism. The large diversity of regulators of apoptosis in mammalian cells and their numerous interactions complicate the analysis of their individual functions, particularly in development. The remarkable conservation of apoptotic mechanisms across species has allowed the genetic pathways of apoptosis determined in lower species, such as the nematode Caenorhabditis elegans and the fruitfly Drosophila melanogaster, to act as models for understanding the biology of apoptosis in mammalian cells. Though many components of the apoptotic pathway are conserved between species, the use of additional model organisms has revealed several important differences and supports the use of model organisms in deciphering complex biological processes such as apoptosis.

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Debcl, a Proapoptotic Bcl-2 Homologue, Is a Component of the Drosophila melanogaster Cell Death Machinery

The Journal of Cell Biology ◽

10.1083/jcb.148.4.703 ◽

2000 ◽

Vol 148 (4) ◽

pp. 703-714 ◽

Cited By ~ 105

Author(s):

Paul A. Colussi ◽

Leonie M. Quinn ◽

David C.S. Huang ◽

Michelle Coombe ◽

Stuart H. Read ◽

...

Keyword(s):

Cell Death ◽

Drosophila Melanogaster ◽

Mammalian Cells ◽

Cultured Cells ◽

Gene Dosage ◽

Family Members ◽

Ectopic Expression ◽

Dependent Manner ◽

Proapoptotic Protein ◽

Main Components

Bcl-2 family of proteins are key regulators of apoptosis. Both proapoptotic and antiapoptotic members of this family are found in mammalian cells, but no such proteins have been described in insects. Here, we report the identification and characterization of Debcl, the first Bcl-2 homologue in Drosophila melanogaster. Structurally, Debcl is similar to Bax-like proapoptotic Bcl-2 family members. Ectopic expression of Debcl in cultured cells and in transgenic flies causes apoptosis, which is inhibited by coexpression of the baculovirus caspase inhibitor P35, indicating that Debcl is a proapoptotic protein that functions in a caspase-dependent manner. debcl expression correlates with developmental cell death in specific Drosophila tissues. We also show that debcl genetically interacts with diap1 and dark, and that debcl-mediated apoptosis is not affected by gene dosage of rpr, hid, and grim. Biochemically, Debcl can interact with several mammalian and viral prosurvival Bcl-2 family members, but not with the proapoptotic members, suggesting that it may regulate apoptosis by antagonizing prosurvival Bcl-2 proteins. RNA interference studies indicate that Debcl is required for developmental apoptosis in Drosophila embryos. These results suggest that the main components of the mammalian apoptosis machinery are conserved in insects.

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BH3-only proteins — evolutionarily conserved proapoptotic Bcl-2 family members essential for initiating programmed cell death

Journal of Cell Science ◽

10.1242/jcs.115.8.1567 ◽

2002 ◽

Vol 115 (8) ◽

pp. 1567-1574 ◽

Cited By ~ 10

Author(s):

Philippe Bouillet ◽

Andreas Strasser

Keyword(s):

Cell Death ◽

Programmed Cell Death ◽

Family Members ◽

Protein Family ◽

Cell Stress ◽

Protective Function ◽

Evolutionarily Conserved ◽

Apoptotic Cascade

The BH3-only members of the Bcl-2 protein family are essential initiators of programmed cell death and are required for apoptosis induced by cytotoxic stimuli. These proteins have evolved to recognise distinct forms of cell stress. In response, they unleash the apoptotic cascade by inactivating the protective function of the pro-survival members of the Bcl-2 family and by activating the Bax/Bax-like pro-apoptotic family members.

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EB1 and EB3 Control CLIP Dissociation from the Ends of Growing Microtubules

Molecular Biology of the Cell ◽

10.1091/mbc.e05-07-0614 ◽

2005 ◽

Vol 16 (11) ◽

pp. 5334-5345 ◽

Cited By ~ 132

Author(s):

Yulia Komarova ◽

Gideon Lansbergen ◽

Niels Galjart ◽

Frank Grosveld ◽

Gary G. Borisy ◽

...

Keyword(s):

Rna Interference ◽

Mammalian Cells ◽

Functional Relationship ◽

Family Members ◽

Microtubule Dynamics ◽

Tyrosine Residue ◽

Dissociation Kinetics ◽

Evolutionarily Conserved ◽

Relationship Of

EBs and CLIPs are evolutionarily conserved proteins, which associate with the tips of growing microtubules, and regulate microtubule dynamics and their interactions with intracellular structures. In this study we investigated the functional relationship of CLIP-170 and CLIP-115 with the three EB family members, EB1, EB2(RP1), and EB3 in mammalian cells. We showed that both CLIPs bind to EB proteins directly. The C-terminal tyrosine residue of EB proteins is important for this interaction. When EB1 and EB3 or all three EBs were significantly depleted using RNA interference, CLIPs accumulated at the MT tips at a reduced level, because CLIP dissociation from the tips was accelerated. Normal CLIP localization was restored by expression of EB1 but not of EB2. An EB1 mutant lacking the C-terminal tail could also fully rescue CLIP dissociation kinetics, but could only partially restore CLIP accumulation at the tips, suggesting that the interaction of CLIPs with the EB tails contributes to CLIP localization. When EB1 was distributed evenly along the microtubules because of overexpression, it slowed down CLIP dissociation but did not abolish its preferential plus-end localization, indicating that CLIPs possess an intrinsic affinity for growing microtubule ends, which is enhanced by an interaction with the EBs.

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Genomic Identification, Evolution, and Expression Analysis of Bromodomain Genes Family in Buffalo

Genes ◽

10.3390/genes13010103 ◽

2022 ◽

Vol 13 (1) ◽

pp. 103

Author(s):

Junjun Zhang ◽

Liangfeng Huang ◽

Pengfei Zhang ◽

Xingchen Huang ◽

Weihan Yang ◽

...

Keyword(s):

Expression Analysis ◽

Developmental Stages ◽

Family Members ◽

Duplication Event ◽

Bubalus Bubalis ◽

Pubertal Stage ◽

Sequencing Platform ◽

Protein Protein Interaction ◽

Structural And Functional Properties ◽

Evolutionarily Conserved

Bromodomain (BRD) is an evolutionarily conserved protein–protein interaction module that is critical in gene regulation, cellular homeostasis, and epigenetics. This study aimed to conduct an identification, evolution, and expression analysis of the BRD gene family in the swamp buffalo (Bubalus bubalis). A total of 101 BRD protein sequences deduced from 22 BRD genes were found in the buffalo genome. The BRD proteins were classified into six groups based on phylogenetic relationships, conserved motifs, and conserved domains. The BRD genes were irregularly distributed in 13 chromosomes. Collinearity analysis revealed 20 BRD gene pairs that had remarkable homologous relationships between the buffalo and cattle, although no tandem or segmental duplication event was found in the buffalo BRD genes. Comparative transcriptomics using a 10x sequencing platform analysis showed that 22 BRD genes were identified in the Sertoli cells (SCs) at different developmental stages of buffalo. Further, the mRNA expression levels of bromodomain and the extraterminal (BET) family in SCs at the pubertal stage were higher than that at the prepubertal stage of buffalo. However, the SMARCA2, PHIP, BRD9, and TAF1 genes exhibited the opposite trend. The maturation process of SCs may be regulated by the BRD family members expressed differentially in SCs at different developmental stages of buffalo. In summary, our findings provide an understanding of the evolutionary, structural, and functional properties of the buffalo BRD family members, and further characterize the function of the BRD family in the maturation of SCs. It also provides a theoretical basis for further understanding in the future of the mechanism of SCs regulating spermatogenesis.

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Comprehensive Analysis of the Prognosis and Correlations With Immune Infiltration of S100 Protein Family Members in Hepatocellular Carcinoma

Frontiers in Genetics ◽

10.3389/fgene.2021.648156 ◽

2021 ◽

Vol 12 ◽

Author(s):

Susu Zheng ◽

Linxia Liu ◽

Tongchun Xue ◽

Chuyu Jing ◽

Xin Xu ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Mutation Rate ◽

S100 Protein ◽

Family Members ◽

Protein Family ◽

Advanced Tumor Stage ◽

Mrna Levels ◽

Expression Levels ◽

Immune Infiltration ◽

Advanced Tumor

S100 protein family members (S100s) are commonly dysregulated in various tumors including hepatocellular carcinoma (HCC). However, the diverse expression, mutation, prognosis and associations with immune infiltration of S100s in HCC have yet to be analyzed. Herein we investigated the roles of S100s in HCC from the Oncomine, Gene Expression Profiling Interactive Analysis (GEPIA), Human Protein Atlas, Kaplan-Meier Plotter, cBioPortal and TIMER databases. Compared with para-cancer tissues, the expression levels of S100A4/S100A6/S100A10/S100A11/S100A13/S100A14/S100P were higher in HCC tissues, while the expression levels of S100A8/S100A9/S100A12 were decreased in tumor tissues. The mRNA levels of S100A2/S100A7/S100A7A/S100A8/S100A9/S100A11 were correlated with advanced tumor stage. Besides, higher mRNA expressions of S100A6/S100A10/S100A11/S100A13/S100A14/S100P were shown to have shorter overall survival (OS), while higher expression of S100A12 was associated with favorable OS. Further, the mutation rate of S100s was investigated, and the high mutation rate (53%) was associated with shorter OS. Additionally, the expressions of S100s were found to be significantly associated with various immune infiltrating cells. Hence, our results showed that S100A6/S100A10/S100A11/S10012/S100A13/S100A14/S100P may be regarded as new prognostic or therapeutic markers and S100s inhibitors may be helpful in the combination of immunotherapies.

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Identification of the Short Neuropeptide F and Short Neuropeptide F Receptor Genes and Their Roles of Food Intake in Dendroctonus armandi

Insects ◽

10.3390/insects12090844 ◽

2021 ◽

Vol 12 (9) ◽

pp. 844

Author(s):

Bin Liu ◽

Danyang Fu ◽

Hang Ning ◽

Ming Tang ◽

Hui Chen

Keyword(s):

Food Intake ◽

Developmental Stages ◽

Control Strategies ◽

Amino Acid Sequences ◽

Body Parts ◽

Expression Levels ◽

Reverse Transcription Pcr ◽

Evolutionarily Conserved ◽

Neuropeptide F ◽

High Degree

The short neuropeptide F (sNPF) is an essential signaling molecule that is evolutionarily conserved and involved in a broad range of physiological functions in the invertebrates, by interacting with sNPF receptors, which belong to G protein-coupled receptors (GPCR). However, the function of sNPF in regulating the food intake of Dendroctonus armandi has been unclear. In this study, we cloned and characterized cDNAs encoding sNPF and sNPF receptor in the D. armandi and made bioinformatics predictions on the deduced amino acid sequences. They had a high degree of similarity to that of Dendroctonus ponderosa. Quantitative real-time reverse transcription PCR (qRT-PCR) revealed that the transcript levels of both sNPF and sNPFR varied across developmental stages and body parts. In addition, the sNPF and sNPFR expression levels were upregulated in starved beetles, and the expression levels recovered after re-feeding. Furthermore, RNAi knockdown by the injection of sNPF and sNPFR dsRNA into beetles significantly increased mortality and reduced their food intake and body weight, and also caused decrease of glycogen and free fatty acid and increase of trehalose. These results indicate that sNPF signaling pathway plays an important role in the regulation of food intake and provides a potential molecular target for the eco-friendly control strategies of this pest.

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Genotype and Trait Specific Responses to Rapamycin Intake in Drosophila melanogaster

Insects ◽

10.3390/insects12050474 ◽

2021 ◽

Vol 12 (5) ◽

pp. 474

Author(s):

Palle Duun Rohde ◽

Asbjørn Bøcker ◽

Caroline Amalie Bastholm Jensen ◽

Anne Louise Bergstrøm ◽

Morten Ib Juul Madsen ◽

...

Keyword(s):

Drosophila Melanogaster ◽

Heat Stress ◽

Side Effects ◽

Protein Kinase ◽

Stress Tolerance ◽

Treated Group ◽

Model System ◽

Treatment Efficiency ◽

Evolutionarily Conserved ◽

Heat Stress Tolerance

Rapamycin is a powerful inhibitor of the TOR (Target of Rapamycin) pathway, which is an evolutionarily conserved protein kinase, that plays a central role in plants and animals. Rapamycin is used globally as an immunosuppressant and as an anti-aging medicine. Despite widespread use, treatment efficiency varies considerably across patients, and little is known about potential side effects. Here we seek to investigate the effects of rapamycin by using Drosophila melanogaster as model system. Six isogenic D. melanogaster lines were assessed for their fecundity, male longevity and male heat stress tolerance with or without rapamycin treatment. The results showed increased longevity and heat stress tolerance for male flies treated with rapamycin. Conversely, the fecundity of rapamycin-exposed individuals was lower than for flies from the non-treated group, suggesting unwanted side effects of the drug in D. melanogaster. We found strong evidence for genotype-by-treatment interactions suggesting that a ‘one size fits all’ approach when it comes to treatment with rapamycin is not recommendable. The beneficial responses to rapamycin exposure for stress tolerance and longevity are in agreement with previous findings, however, the unexpected effects on reproduction are worrying and need further investigation and question common believes that rapamycin constitutes a harmless drug.

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