Limits...
ZmbZIP60 mRNA is spliced in maize in response to ER stress.

Li Y, Humbert S, Howell SH - BMC Res Notes (2012)

Bottom Line: Splicing converts the predicted protein from a membrane-associated transcription factor to one that is targeted to the nucleus.Maize has numerous BIP-like genes, and ER stress was found to upregulate one of these, ZmBIPb.Among the genes upregulated by ER stress in maize is one of 22 BIP-like genes, ZmBIPb.

View Article: PubMed Central - HTML - PubMed

Affiliation: Plant Sciences Institute, Iowa State University, Ames, IA 50014, USA.

ABSTRACT

Background: Adverse environmental conditions produce ER stress and elicit the unfolded protein response (UPR) in plants. Plants are reported to have two "arms" of the ER stress signaling pathway-one arm involving membrane-bound transcription factors and the other involving a membrane-associated RNA splicing factor, IRE1. IRE1 in yeast to mammals recognizes a conserved twin loop structure in the target RNA.

Results: A segment of the mRNA encoding ZmbZIP60 in maize can be folded into a twin loop structure, and in response to ER stress this mRNA is spliced, excising a 20b intron. Splicing converts the predicted protein from a membrane-associated transcription factor to one that is targeted to the nucleus. Splicing of ZmbZIP60 can be elicited in maize seedlings by ER stress agents such as dithiothreitol (DTT) or tunicamycin (TM) or by heat treatment. Younger, rather than older seedlings display a more robust splicing response as do younger parts of leaf, along a developmental gradient in a leaf. The molecular signature of an ER stress response in plants includes the upregulation of Binding Protein (BIP) genes. Maize has numerous BIP-like genes, and ER stress was found to upregulate one of these, ZmBIPb.

Conclusions: The splicing of ZmbZIP60 mRNA is an indicator of ER stress in maize seedlings resulting from adverse environmental conditions such as heat stress. ZmbZIP60 mRNA splicing in maize leads predictively to the formation of active bZIP transcription factor targeted to the nucleus to upregulate stress response genes. Among the genes upregulated by ER stress in maize is one of 22 BIP-like genes, ZmBIPb.

Show MeSH

Related in: MedlinePlus

PCR assays used in this study. (A) Flanking primer (FP) assay employs primers immediately flanking the splice site, so PCR products from both the unspliced and spliced Zmbzip60 are amplified. (B) Specific primer unspliced (SPU) assay uses a primer that crosses the exon-intron boundary and is specific for detecting the unspliced form of bzip60. Specific primers spliced (SPS) assay uses a primer that crosses the exon-exon boundary and is specific for detecting the spliced form of ZmbZIP60. The specificity of the two SP assays was demonstrated by amplifying the plasmid-cloned cDNA derived from the spliced and unspliced bzip60.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3369818&req=5

Figure 2: PCR assays used in this study. (A) Flanking primer (FP) assay employs primers immediately flanking the splice site, so PCR products from both the unspliced and spliced Zmbzip60 are amplified. (B) Specific primer unspliced (SPU) assay uses a primer that crosses the exon-intron boundary and is specific for detecting the unspliced form of bzip60. Specific primers spliced (SPS) assay uses a primer that crosses the exon-exon boundary and is specific for detecting the spliced form of ZmbZIP60. The specificity of the two SP assays was demonstrated by amplifying the plasmid-cloned cDNA derived from the spliced and unspliced bzip60.

Mentions: To determine if ZmbZIP60 is spliced in a manner similar to AtbZIP60, we exposed corn seedlings to ER stress agents such as dithiothreitol (DTT) and tested for ZmbZIP60 splicing using RT-PCR assays. Two assays were used as described before [27]-one, a flanking primers (FP) assay, which detects both the unspliced and spliced bZIP60 mRNAs (Figure 2A), and another, a specific primers (SP) assay, which detects either the spliced (SPS) or the unspliced (SPU) forms of bZIP60 (Figure 2B).


ZmbZIP60 mRNA is spliced in maize in response to ER stress.

Li Y, Humbert S, Howell SH - BMC Res Notes (2012)

PCR assays used in this study. (A) Flanking primer (FP) assay employs primers immediately flanking the splice site, so PCR products from both the unspliced and spliced Zmbzip60 are amplified. (B) Specific primer unspliced (SPU) assay uses a primer that crosses the exon-intron boundary and is specific for detecting the unspliced form of bzip60. Specific primers spliced (SPS) assay uses a primer that crosses the exon-exon boundary and is specific for detecting the spliced form of ZmbZIP60. The specificity of the two SP assays was demonstrated by amplifying the plasmid-cloned cDNA derived from the spliced and unspliced bzip60.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3369818&req=5

Figure 2: PCR assays used in this study. (A) Flanking primer (FP) assay employs primers immediately flanking the splice site, so PCR products from both the unspliced and spliced Zmbzip60 are amplified. (B) Specific primer unspliced (SPU) assay uses a primer that crosses the exon-intron boundary and is specific for detecting the unspliced form of bzip60. Specific primers spliced (SPS) assay uses a primer that crosses the exon-exon boundary and is specific for detecting the spliced form of ZmbZIP60. The specificity of the two SP assays was demonstrated by amplifying the plasmid-cloned cDNA derived from the spliced and unspliced bzip60.
Mentions: To determine if ZmbZIP60 is spliced in a manner similar to AtbZIP60, we exposed corn seedlings to ER stress agents such as dithiothreitol (DTT) and tested for ZmbZIP60 splicing using RT-PCR assays. Two assays were used as described before [27]-one, a flanking primers (FP) assay, which detects both the unspliced and spliced bZIP60 mRNAs (Figure 2A), and another, a specific primers (SP) assay, which detects either the spliced (SPS) or the unspliced (SPU) forms of bZIP60 (Figure 2B).

Bottom Line: Splicing converts the predicted protein from a membrane-associated transcription factor to one that is targeted to the nucleus.Maize has numerous BIP-like genes, and ER stress was found to upregulate one of these, ZmBIPb.Among the genes upregulated by ER stress in maize is one of 22 BIP-like genes, ZmBIPb.

View Article: PubMed Central - HTML - PubMed

Affiliation: Plant Sciences Institute, Iowa State University, Ames, IA 50014, USA.

ABSTRACT

Background: Adverse environmental conditions produce ER stress and elicit the unfolded protein response (UPR) in plants. Plants are reported to have two "arms" of the ER stress signaling pathway-one arm involving membrane-bound transcription factors and the other involving a membrane-associated RNA splicing factor, IRE1. IRE1 in yeast to mammals recognizes a conserved twin loop structure in the target RNA.

Results: A segment of the mRNA encoding ZmbZIP60 in maize can be folded into a twin loop structure, and in response to ER stress this mRNA is spliced, excising a 20b intron. Splicing converts the predicted protein from a membrane-associated transcription factor to one that is targeted to the nucleus. Splicing of ZmbZIP60 can be elicited in maize seedlings by ER stress agents such as dithiothreitol (DTT) or tunicamycin (TM) or by heat treatment. Younger, rather than older seedlings display a more robust splicing response as do younger parts of leaf, along a developmental gradient in a leaf. The molecular signature of an ER stress response in plants includes the upregulation of Binding Protein (BIP) genes. Maize has numerous BIP-like genes, and ER stress was found to upregulate one of these, ZmBIPb.

Conclusions: The splicing of ZmbZIP60 mRNA is an indicator of ER stress in maize seedlings resulting from adverse environmental conditions such as heat stress. ZmbZIP60 mRNA splicing in maize leads predictively to the formation of active bZIP transcription factor targeted to the nucleus to upregulate stress response genes. Among the genes upregulated by ER stress in maize is one of 22 BIP-like genes, ZmBIPb.

Show MeSH
Related in: MedlinePlus