Identification of two cis-acting elements that independently regulate the length of poly(A) on Xenopus albumin pre-mRNA

Gupta, Jaydip Das; Gu, Howard H.; Chernokalskaya, Elena; Gao, Xiang; Schoenberg, Daniel R.

doi:10.1017/s1355838298971837

Cited by 23 publications

(30 citation statements)

References 34 publications

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“…If this is eventually confirmed, then consideration of a separate pathway for efficient nuclear export must be considered, since the 3Ј-UTR CARE did not affect cytoplasmic mRNA levels. This activity parallels that reported for the poly(A)-limiting element, which reduced poly(A) tail length in the nucleus without affecting nuclear export or polysomal loading (38,39). The observation that the 3Ј-UTR CARE limits translation without altering polysomal loading is similar to that seen with other systems (33,34).…”

Section: Post-transcriptional Regulation Of Cd154 Expression Bysupporting

confidence: 86%

Separate cis-trans Pathways Post-transcriptionally Regulate Murine CD154 (CD40 Ligand) Expression

Hamilton

Wang

Collins

et al. 2008

Journal of Biological Chemistry

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We report a role for CA repeats in the 3 -untranslated region (3 -UTR) in regulating CD154 expression. Human CD154 is encoded by an unstable mRNA; this instability is conferred in cis by a portion of its 3 -UTR that includes a polypyrimidine-rich region and CA dinucleotide repeat. We demonstrate similar instability activity with the murine CD154 3 -UTR. This instability element mapped solely to a conserved 100-base CU-rich region alone, which we call a CU-rich response element. Surprisingly, the CA dinucleotide-rich region also regulated reporter expression but at the level of translation. This activity was associated with poly(A) tail shortening and regulated by heterogeneous nuclear ribonucleoprotein L levels. We conclude that the CD154 3 -UTR contains dual cis-acting elements, one of which defines a novel function for exonic CA dinucleotide repeats. These findings suggest a mechanism for the association of 3 -UTR CA-rich response element polymorphisms with CD154 overexpression and the subsequent risk of autoimmune disease.

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Section: Post-transcriptional Regulation Of Cd154 Expression Bysupporting

confidence: 86%

Separate cis-trans Pathways Post-transcriptionally Regulate Murine CD154 (CD40 Ligand) Expression

Hamilton

Wang

Collins

et al. 2008

Journal of Biological Chemistry

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“…In the experiment in Figure 1, LM(tkÀ) cells were transfected with plasmid vectors CMV-glo-SPA (control), which produces b-globin mRNA with a long, heterogeneous 100-to 200-nt poly(A) tail, or CMV-glo-PLE B-SPA (+PLE), which carries the conserved PLE B element (Gu et al 1999), and produces b-globin mRNA with a discrete, <20-nt poly(A) tail (Das Gupta et al 1998). A plasmid expressing firefly luciferase was included as a cotransfected control.…”

Section: Resultsmentioning

confidence: 99%

“…The construction of the plasmids CMV-glo-SPA and CMV-glo-PLE B-SPA and the inactive MutG form of the PLE were described previously (Das Gupta et al 1998). Briefly, a plasmid bearing the human b-globin gene under control of the CMV promoter was cut in exon 3 (position 1497) by EcoRI.…”

Section: Plasmid Constructsmentioning

confidence: 99%

“…The poly(A) tail is shortened at various rates during the lifetime of the mRNA, ultimately leading to deadenylation followed by degradation of the mRNA body (Wilusz et al 2001). We previously showed that Xenopus albumin mRNA has a <20-nt poly(A) tail (Schoenberg et al 1989;Rao et al 1996) and identified a unique element, the poly(A)-limiting element, or PLE, as the cis-acting sequence responsible for this short poly(A) phenotype (Das Gupta et al 1998). PLEs have also been confirmed in transferrin mRNA and the mRNA encoding the transcription factor HIV-EP2 (Gu et al 1999), and the 3 0 ends of a number of genes contain similar sequence elements.…”

Section: Introductionmentioning

confidence: 99%

“…Both fully processed cytoplasmic albumin mRNA and unspliced albumin pre-mRNA have <20-nt poly(A) tails, indicating that the restriction in poly(A) length occurs on nuclear pre-mRNA (Rao et al 1996). Deletion analysis identified two redundant PLEs in the terminal exon of albumin pre-mRNA (Das Gupta et al 1998), either of which changed the length of the poly(A) tail on a human b-globin reporter mRNA from $200 nt to <20 nt when placed upstream of a highly efficient synthetic polyadenylation element (SPA) (Levitt et al 1989;Das Gupta et al 1998). To function in regulating poly(A) length the PLE must be in the terminal exon, and as long this location is maintained the distance between it and AAUAAA could be changed without affecting its ability to restrict poly(A) to <20 nt (Das Gupta et al 2001).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The poly(A)-limiting element enhances mRNA accumulation by increasing the efficiency of pre-mRNA 3′ processing

Peng

Murray²,

Schoenberg³

2005

RNA

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The poly(A)-limiting element (PLE) is a conserved sequence originally found in the 3 0 UTR of Xenopus albumin mRNA whose presence restricts the length of the poly(A) tail on both pre-mRNA and fully processed mRNA to <20 nt. Results presented in this study show that the PLE also increases the cytoplasmic level of reporter b-globin mRNA. Transcription run-on shows this increase was not due to increased reporter gene transcription, and experiments with tetracycline repressor-controlled reporter mRNA showed the PLE does not alter the rate of mRNA decay. Both RT-PCR and RNase protection assay showed the PLE caused a 50% increase in the 3 0 processing of reporter b-globin mRNA in vivo. This was confirmed in vitro, where PLE-containing RNA was cleaved in HeLa nuclear extract at a rate 80% faster than a control RNA bearing an inactive element. These results indicate that the PLE regulates the length of the poly(A) tail and the efficiency of 3 0 processing. In addition, they show that PLE-containing mRNA with a <20-nt poly(A) tail is as stable as mRNA with a 100-to 200-nt poly(A) tail.

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Measuring the tail: Methods for poly(A) tail profiling

et al. 2022

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The 3 0 -end poly(A) tail is an important and potent feature of most mRNA molecules that affects mRNA fate and translation efficiency. Polyadenylation is a posttranscriptional process that occurs in the nucleus by canonical poly(A) polymerases (PAPs). In some specific instances, the poly(A) tail can also be extended in the cytoplasm by noncanonical poly(A) polymerases (ncPAPs). This epitranscriptomic regulation of mRNA recently became one of the most interesting aspects in the field. Advances in RNA sequencing technologies and software development have allowed the precise measurement of poly(A) tails, identification of new ncPAPs, expansion of the function of known enzymes, discovery and a better understanding of the physiological role of tail heterogeneity, and recognition of a correlation between tail length and RNA translatability. Here, we summarize the development of polyadenylation research methods, including classic low-throughput approaches, Illumina-based genomewide analysis, and advanced state-of-art techniques that utilize long-read third-generation sequencing with Pacific Biosciences and Oxford Nanopore Technologies platforms. A boost in technical opportunities over recent decades has allowed a better understanding of the regulation of gene expression at the mRNA level. This article is categorized under: RNA Methods > RNA Analyses In Vitro and In Silico K E Y W O R D S 3 0 -end tailing, poly(A) tail, regulation of gene expression, RNA, RNA sequencing

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Identification of two cis-acting elements that independently regulate the length of poly(A) on Xenopus albumin pre-mRNA

Cited by 23 publications

References 34 publications

Separate cis-trans Pathways Post-transcriptionally Regulate Murine CD154 (CD40 Ligand) Expression

Separate cis-trans Pathways Post-transcriptionally Regulate Murine CD154 (CD40 Ligand) Expression

The poly(A)-limiting element enhances mRNA accumulation by increasing the efficiency of pre-mRNA 3′ processing

Measuring the tail: Methods for poly(A) tail profiling

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