Diverse soybean actin transcripts contain a large intron in the 5? untranslated leader: structural similarity to vertebrate muscle actin genes

Pearson, Leslie; Meagher, Richard B.

doi:10.1007/bf00027497

Cited by 26 publications

(21 citation statements)

References 63 publications

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“…The ACT8 transcript had multiple 5' ends at 33, 37, and 42 nt downstream of the TATATA sequence. Multiple transcription start sites have been identified in several plant actin genes (An et al, 1996;Pearson and Meagher, 1990). Alignment of the 5' flanking sequences (Figure 2) revealed a 9 bp deletion/insertion difference between the TATA boxes and the transcriptional start sites of ACT2 and ACT8.…”

Section: Isolation Of the Act2 And Act8 Actin Genes And Characterizatmentioning

confidence: 99%

Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissues

et al. 1996

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SummaryArabidopsis has a complex and ancient actin gene family encoding six divergent subclasses of proteins. One subclass is represented by ACT2 and ACT8, which encode nearly identical proteins. These two genes differ significantly in flanking and intron sequences and in silent nucleotide positions within codons. Gene-specific RNA gel blot hybridization and reverse transcriptase-mediated polymerase chain reaction (RT-PCR) assays showed that ACT2 and/or ACT8 mRNAs were coordinately and strongly expressed in leaves, roots, stems, flowers, pollen, and siliques. Together they account for greater than 80% of the actin mRNA in most Arabidopsis organs. The 5' flanking regions, including the promoter, the mRNA leader exon, an intron in the mRNA leader, and the first 19 codons, were coupled to a 13-glucuronidase (GUS) reporter gene and transformed into Arabidopsis. The ACT2/GUS construct was expressed strongly in nearly all the vegetative tissues in seedlings, juvenile plants, and mature plants. These activities persisted in older tissues. Little or no expression was observed in seed coats, hypocotyls, gynoecia, or pollen sacs. In contrast, the expression of the ACT8/GUS construct was weaker. It was observed only in a subset of the organs and tissues expressing ACT2/GUS and was not significantly expressed in the flower. ACT2, ACT8, and ACT8/GUS mRNAs were present at moderate to high levels in pollen, and yet neither ACT2/GUS nor ACT8/GUS enzyme expression could be detected in pollen.This suggested a mechanism of translational control affecting ACT2 and ACT8 expression in some tissues. The conservation of protein sequence and overlapping patterns of expression, in spite of significant DNA sequence divergence, suggests that the function and regulation of these two genes have been conserved during the evolution of the Brassicaceae.

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Section: Isolation Of the Act2 And Act8 Actin Genes And Characterizatmentioning

confidence: 99%

Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissues

et al. 1996

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“…These locations are identical to those in most characterized plant actin genes (Meagher and Williamson, 1994). A few well-characterized plant actin genes have been shown to contain a large intron in the 5' untranslated region (5' UTR) of the mRNA (McElroy et al, 1990b;Pearson and Meagher, 1990). To determine whether ACT7 and ACT3 have an intron in this position, the 5' UTRs of their cDNAs were amplified from a flower cDNA library by a polymerase chain reaction (PCR) protocol (see details in Methods).…”

Section: Lsolation and Characterization Of Ac7y And Act3mentioning

confidence: 99%

“…The TATA box, located 37 nucleotides upstream of this site, is apparently conserved between ACT1 and ACT3. Multiple start sites have been found in many of the plant actin transcripts that have been examined in detail (Pearson and Meagher, 1990).…”

Section: Lsolation and Characterization Of Ac7y And Act3mentioning

confidence: 99%

“…Large introns interrupting the mRNA leader are not common among plant genes but are found in actin genes from rice, tobacco, soybean, and vertebrates (Nakajima-lijima et al, 1985;McElroy et al, 1990b;Pearson and Meagher, 1990;Thangavelu et al, 1993). McElroy et al (1990b) reported that the intron in the 5'UTR of rice actin, RAc7, is essential for strong expression in electroporated rice protoplasts.…”

Section: Potential Elements Affecting Ac77 Act3 and 7737 Regulationmentioning

confidence: 99%

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Conserved expression of the Arabidopsis ACT1 and ACT 3 actin subclass in organ primordia and mature pollen.

Huang

McDowell

et al. 1996

Plant Cell

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We have proposed that ancient and divergent classes of plant actin genes have been preserved throughout vascular plant evolution, because they have distinct patterns of gene regulation. The hypothesis was explored for AC7Y and ACT3, which represent one of the six ancient subclasses in the Arabidopsis actin gene family. Comparison of AC7Y and ACT3 cDNA and genomic sequences revealed highly divergent flanking and intron sequences, whereas they encoded nearly identical proteins. Quantification of their leve1 of divergence suggests that they have not shared a common ancestor for 30 to 60 million years. Gene-specific FINA gel blot hybridization and reverse transcriptase-polymerase chain reaction analyses demonstrated that the distribution of AC7Y and ACT3 mRNAs was very similar: both preferentially accumulated at high levels in mature pollen and at very low levels in the other major organs. The 5' flanking regions of both genes, including the promoter, leader exon and intron, and thefirst 19 codons, werefused to the P-giucuronidase (GUS) reporter gene. The expression of these reporter fusions was examined in a large number of transgenic Arabidopsis plants. Histochemical assays demonstrated that both ACTT-GUS and ACT3-GUS constructs were expressed preferentially in pollen, pollen tubes, and in all organ primordia, including those in roots, shoots, and the inflorescence. Comparison of the 5' flanking regions of AC7Y and ACT3 revealed a number of short conserved sequences, which may direct their common transcriptional and post-transcriptional regulation. The expression patterns observed were distinct from those of any other Arabidopsis actin subclass. The conservation of their expression pattern and amino acid sequences suggests that this actin subclass plays a distinct and required role in the plant cytoskeleton.

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“…After (19,23,24,28,43,(46)(47)(48). These elements have also been found in the 5' regions of several plant actin genes (38,39). It has been proposed that polypurine/polypyrimidine elements affect local chromatin structure by changing nucleosome arrangement, thus facilitating access of transcription factors to the promoter DNA (15,22).…”

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confidence: 99%

Characterization of cis-acting elements regulating transcription from the promoter of a constitutively active rice actin gene.

et al. 1992

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The promoter of the constitutively expressed rice (Oryza sativa) actin 1 gene (Act1) is highly active in transformed rice plants (W. Zhang, D. McElroy, and R. Wu, Plant Cell 3:1150-1160, 1991). A region 834 bp upstream of the Act1 transcription initiation site contains all the regulatory elements necessary for maximal gene expression in transformed rice protoplasts (D. McElroy, W. Zhang, J. Cao, and R. Wu, Plant Cell 2:163-171, 1990). We have constructed a series of Act1 promoter deletions fused to a bacterial beta-glucuronidase reporter sequence (Gus). Transient expression assays in transformed rice protoplasts, as well as transformed maize cells and tissues, identified two distinct cis-acting regulatory elements in the Act1 promoter. A 38-bp poly(dA-dT) region was found to be a positive regulator of Act1 promoter activity. Deletion of the poly(dA-dT) element lowered Gus expression by at least threefold compared with expression produced by the full-length Act1 promoter. By gel retardation and footprinting, we identified a ubiquitous rice protein which specifically recognizes this poly(dA-dT) element in the constitutively active Act1 promoter. A CCCAA pentamer repeat-containing region was found to be a negative regulator of the Act1 promoter in transformed rice protoplasts. Transient expression assays in different maize cells and tissues with use of the Act1 deletion constructs suggested that the CCCAA pentamer repeat region functions in a complex tissue-specific manner. A CCCAA-binding protein was detected only in root extracts.

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Diverse soybean actin transcripts contain a large intron in the 5? untranslated leader: structural similarity to vertebrate muscle actin genes

Cited by 26 publications

References 63 publications

Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissues

Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissues

Conserved expression of the Arabidopsis ACT1 and ACT 3 actin subclass in organ primordia and mature pollen.

Characterization of cis-acting elements regulating transcription from the promoter of a constitutively active rice actin gene.

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