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Bombyx E75 isoforms display stage- and tissue-specific responses to 20-hydroxyecdysone.

Li K, Guo E, Hossain MS, Li Q, Cao Y, Tian L, Deng X, Li S - Sci Rep (2015)

Bottom Line: At the early 5(th) larval instar stage, treatment of the prothoracic glands and fat body with 20-hydroxyecdysone (20E) and/or cycloheximide (CHX) revealed that BmE75A is 20E primary-responsive at both mRNA and protein levels, while BmE75B and BmE75C exhibit various responses to 20E.At the early wandering stage, RNAi-mediated reduction of gene expression of the 20E nuclear receptor complex, EcR-USP, significantly decreased mRNA and protein levels of all three BmE75 isoforms in both tissues.In conclusion, BmE75 isoforms display stage- and tissue-specific responses to 20E at both mRNA and protein levels; moreover, they are regulated by other unknown factors at the protein level.

View Article: PubMed Central - PubMed

Affiliation: 1] Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/Guangdong Provincial Sericulture and Mulberry Engineering Research Center, College of Animal Sciences, South China Agricultural University, Guangzhou, 510642, China [2] Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.

ABSTRACT
Resulted from alternative splicing of the 5' exons, the nuclear receptor gene E75 in the silkworm, Bombyx mori, processes three mRNA isoforms, BmE75A, BmE75B and BmE75C. From the early 5(th) larval instar to the prepupal stages, BmE75A mRNA and protein levels in the prothoracic glands display developmental profiles similar to ecdysteroid titer. In the fat body, mRNA levels but not protein levels of all three BmE75 isoforms correlate with ecdysteroid titer; moreover, proteins of all three BmE75 isoforms disappear at the prepupal stages, and a modified BmE75 protein with smaller molecular weight and cytoplasm localization occurs. At the early 5(th) larval instar stage, treatment of the prothoracic glands and fat body with 20-hydroxyecdysone (20E) and/or cycloheximide (CHX) revealed that BmE75A is 20E primary-responsive at both mRNA and protein levels, while BmE75B and BmE75C exhibit various responses to 20E. At the early wandering stage, RNAi-mediated reduction of gene expression of the 20E nuclear receptor complex, EcR-USP, significantly decreased mRNA and protein levels of all three BmE75 isoforms in both tissues. In conclusion, BmE75 isoforms display stage- and tissue-specific responses to 20E at both mRNA and protein levels; moreover, they are regulated by other unknown factors at the protein level.

No MeSH data available.


Three BmE75 isoforms.(A) Genomic location of the three transcripts of BmE75. Gray boxes denote common 3′ exons. Purple, yellow and green boxes denote specific 5′ exons of BmE75A, BmE75B and BmE75C, respectively. Diagram of BmE75A, BmE75B and BmE75C protein structures. Different colors denote protein domains correspond to the product of exons in (A). DBD, DNA binding domon; LBD, ligand binding domon; AF-1, activitiaon function-1; AF-2, activitiaon function-2. The red line shows the region to generate the BmE75 antibody.
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f1: Three BmE75 isoforms.(A) Genomic location of the three transcripts of BmE75. Gray boxes denote common 3′ exons. Purple, yellow and green boxes denote specific 5′ exons of BmE75A, BmE75B and BmE75C, respectively. Diagram of BmE75A, BmE75B and BmE75C protein structures. Different colors denote protein domains correspond to the product of exons in (A). DBD, DNA binding domon; LBD, ligand binding domon; AF-1, activitiaon function-1; AF-2, activitiaon function-2. The red line shows the region to generate the BmE75 antibody.

Mentions: As shown in SilkDB (http://silkworm.genomics.org.cn/silkdb/), the BmE75 gene locus is located on chromosome 10, spanning 130 kb of genomic DNA. Each BmE75 isoform is characterized by a unique N-terminal sequence encoded by one (BmE75A and BmE75B) or two (BmE75C) distinct 5′ exons. These 5′ exons splice to a common set of four 3′ exons for BmE75A and BmE75C, while BmE75B shares only the last three 3′ exons (Fig. 1A). Overall, the three BmE75 isoforms, BmE75A, BmE75B and BmE75C, originate from a single copy gene through differential promoter usage and alternative splicing of 5′ exons, showing similar arrangement of gene structure to DmE757141516.


Bombyx E75 isoforms display stage- and tissue-specific responses to 20-hydroxyecdysone.

Li K, Guo E, Hossain MS, Li Q, Cao Y, Tian L, Deng X, Li S - Sci Rep (2015)

Three BmE75 isoforms.(A) Genomic location of the three transcripts of BmE75. Gray boxes denote common 3′ exons. Purple, yellow and green boxes denote specific 5′ exons of BmE75A, BmE75B and BmE75C, respectively. Diagram of BmE75A, BmE75B and BmE75C protein structures. Different colors denote protein domains correspond to the product of exons in (A). DBD, DNA binding domon; LBD, ligand binding domon; AF-1, activitiaon function-1; AF-2, activitiaon function-2. The red line shows the region to generate the BmE75 antibody.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499807&req=5

f1: Three BmE75 isoforms.(A) Genomic location of the three transcripts of BmE75. Gray boxes denote common 3′ exons. Purple, yellow and green boxes denote specific 5′ exons of BmE75A, BmE75B and BmE75C, respectively. Diagram of BmE75A, BmE75B and BmE75C protein structures. Different colors denote protein domains correspond to the product of exons in (A). DBD, DNA binding domon; LBD, ligand binding domon; AF-1, activitiaon function-1; AF-2, activitiaon function-2. The red line shows the region to generate the BmE75 antibody.
Mentions: As shown in SilkDB (http://silkworm.genomics.org.cn/silkdb/), the BmE75 gene locus is located on chromosome 10, spanning 130 kb of genomic DNA. Each BmE75 isoform is characterized by a unique N-terminal sequence encoded by one (BmE75A and BmE75B) or two (BmE75C) distinct 5′ exons. These 5′ exons splice to a common set of four 3′ exons for BmE75A and BmE75C, while BmE75B shares only the last three 3′ exons (Fig. 1A). Overall, the three BmE75 isoforms, BmE75A, BmE75B and BmE75C, originate from a single copy gene through differential promoter usage and alternative splicing of 5′ exons, showing similar arrangement of gene structure to DmE757141516.

Bottom Line: At the early 5(th) larval instar stage, treatment of the prothoracic glands and fat body with 20-hydroxyecdysone (20E) and/or cycloheximide (CHX) revealed that BmE75A is 20E primary-responsive at both mRNA and protein levels, while BmE75B and BmE75C exhibit various responses to 20E.At the early wandering stage, RNAi-mediated reduction of gene expression of the 20E nuclear receptor complex, EcR-USP, significantly decreased mRNA and protein levels of all three BmE75 isoforms in both tissues.In conclusion, BmE75 isoforms display stage- and tissue-specific responses to 20E at both mRNA and protein levels; moreover, they are regulated by other unknown factors at the protein level.

View Article: PubMed Central - PubMed

Affiliation: 1] Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/Guangdong Provincial Sericulture and Mulberry Engineering Research Center, College of Animal Sciences, South China Agricultural University, Guangzhou, 510642, China [2] Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.

ABSTRACT
Resulted from alternative splicing of the 5' exons, the nuclear receptor gene E75 in the silkworm, Bombyx mori, processes three mRNA isoforms, BmE75A, BmE75B and BmE75C. From the early 5(th) larval instar to the prepupal stages, BmE75A mRNA and protein levels in the prothoracic glands display developmental profiles similar to ecdysteroid titer. In the fat body, mRNA levels but not protein levels of all three BmE75 isoforms correlate with ecdysteroid titer; moreover, proteins of all three BmE75 isoforms disappear at the prepupal stages, and a modified BmE75 protein with smaller molecular weight and cytoplasm localization occurs. At the early 5(th) larval instar stage, treatment of the prothoracic glands and fat body with 20-hydroxyecdysone (20E) and/or cycloheximide (CHX) revealed that BmE75A is 20E primary-responsive at both mRNA and protein levels, while BmE75B and BmE75C exhibit various responses to 20E. At the early wandering stage, RNAi-mediated reduction of gene expression of the 20E nuclear receptor complex, EcR-USP, significantly decreased mRNA and protein levels of all three BmE75 isoforms in both tissues. In conclusion, BmE75 isoforms display stage- and tissue-specific responses to 20E at both mRNA and protein levels; moreover, they are regulated by other unknown factors at the protein level.

No MeSH data available.