The Association of MEG3 lncRNA with Nuclear Speckles in Living Cells

Hasenson, Sarah E; Alkalay, Ella; Atrash, Mohammad K; Boocholez, Alon; Gershbaum, Julianna; Hochberg-Laufer, Hodaya; Shav‐Tal, Yaron

doi:10.3390/cells11121942

Cited by 6 publications

(4 citation statements)

References 71 publications

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“…To address this issue, we applied the commonly used splicing inhibitor PLB, which inhibits SF3B1, an important component in complex A of the spliceosome ( 38 ). Treatment with PLB for 6 h led to an enlargement in the size of nuclear speckles ( 39 ), a known indication for inhibition of pre-mRNA splicing. In addition, this treatment also resulted in reduced size of arsenite-induced SGs, compared to cells treated with arsenite only (Figure 2A and B).…”

Section: Resultsmentioning

confidence: 99%

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Nuclear RNA-related processes modulate the assembly of cytoplasmic RNA granules

Angel,

Fleshler,

Atrash

et al. 2024

Nucleic Acids Research

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Stress granules (SGs) are cytoplasmic assemblies formed under various stress conditions as a consequence of translation arrest. SGs contain RNA-binding proteins, ribosomal subunits and messenger RNAs (mRNAs). It is well known that mRNAs contribute to SG formation; however, the connection between SG assembly and nuclear processes that involve mRNAs is not well established. Here, we examine the effects of inhibiting mRNA transcription, splicing and export on the assembly of SGs and the related cytoplasmic P body (PB). We demonstrate that inhibition of mRNA transcription, splicing and export reduces the formation of canonical SGs in a eukaryotic initiation factor 2α phosphorylation-independent manner, and alters PB size and quantity. We find that the splicing inhibitor madrasin promotes the assembly of stress-like granules. We show that the addition of synthetic mRNAs directly to the cytoplasm is sufficient for SG assembly, and that the assembly of these SGs requires the activation of stress-associated protein synthesis pathways. Moreover, we show that adding an excess of mRNA to cells that do not have active splicing, and therefore have low levels of cytoplasmic mRNAs, promotes SG formation under stress conditions. These findings emphasize the importance of the cytoplasmic abundance of newly transcribed mRNAs in the assembly of SGs.

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Section: Resultsmentioning

confidence: 99%

“…It is known that splicing inhibition leads to an enlargement of nuclear speckles ( 51 ). The observation that different splicing inhibitors affect PBs differently led us to question whether nuclear speckles would be enlarged in cells treated with the other splicing inhibitors.…”

Section: Resultsmentioning

confidence: 99%

Nuclear RNA-related processes modulate the assembly of cytoplasmic RNA granules

Angel,

Fleshler,

Atrash

et al. 2024

Nucleic Acids Research

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“…This is a rare occurrence, since introns are non-coding sequences that do not reach the cytoplasm. Moreover, when splicing is inhibited, the unspliced transcripts are usually retained at the site of transcription or accumulate in nuclear speckles (Hasenson et al 2022 ; Barutcu et al 2022 ; Hall et al 2006 ; Johnson et al 2000 ; Mor et al 2016 ). These transcripts do not exit the nucleus since they are detected as aberrant transcripts.…”

Section: Discussionmentioning

confidence: 99%

CCAT1 lncRNA is chromatin-retained and post-transcriptionally spliced

Bohrer,

Varon,

Peretz

et al. 2024

Histochem Cell Biol

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Super-enhancers are unique gene expression regulators widely involved in cancer development. Spread over large DNA segments, they tend to be found next to oncogenes. The super-enhancer c-MYC locus forms long-range chromatin looping with nearby genes, which brings the enhancer and the genes into proximity, to promote gene activation. The colon cancer-associated transcript 1 (CCAT1) gene, which is part of the MYC locus, transcribes a lncRNA that is overexpressed in colon cancer cells through activation by MYC. Comparing different types of cancer cell lines using RNA fluorescence in situ hybridization (RNA FISH), we detected very prominent CCAT1 expression in HeLa cells, observed as several large CCAT1 nuclear foci. We found that dozens of CCAT1 transcripts accumulate on the gene locus, in addition to active transcription occurring from the gene. The accumulating transcripts are released from the chromatin during cell division. Examination of CCAT1 lncRNA expression patterns on the single-RNA level showed that unspliced CCAT1 transcripts are released from the gene into the nucleoplasm. Most of these unspliced transcripts were observed in proximity to the active gene but were not associated with nuclear speckles in which unspliced RNAs usually accumulate. At larger distances from the gene, the CCAT1 transcripts appeared spliced, implying that most CCAT1 transcripts undergo post-transcriptional splicing in the zone of the active gene. Finally, we show that unspliced CCAT1 transcripts can be detected in the cytoplasm during splicing inhibition, which suggests that there are several CCAT1 variants, spliced and unspliced, that the cell can recognize as suitable for export.

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“…Long non-coding RNA (lncRNA) is a class of RNA molecules that are widely present in cells (Ahmadpour ST et al, 2023;Hasenson SE et al, 2022). They are usually longer than 200 nucleotides and do unable to transform proteins (Nojima T et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

Long noncoding RNA LINC01842 enhances disulfidptosis resistance and promotes glioma progression via IQGAP1

Zhang,

Wang,

Zheng

et al. 2024

Preprint

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Background: Long non-coding RNA (lncRNA) is widely present in cells and is demonstrated to play vital roles in the development and progression of glioma. However, the biological roles and function mechanisms of LINC01842 in glioma are not yet clear. This study aims to elucidate the potential role of LINC01842 in glioma, specifically its association with disulfidptosis. Methods: We obtained sequencing data from The Cancer Gene Atlas (TCGA). Correction analysis was applied to select disulfidptosis-related lncRNAs (DRLs). Prognosis-associated DRLs were identified by least absolute shrinkage and selection operator (LASSO) and support vector machine (SVM). Receiver operating characteristic (ROC) curve and multivariate Cox regression analyses were used to screen our target gene LINC01842. Subsequently, expression of LINC01842, IQGAP1and SLC7A11 in glioma cell lines was detected by real-time quantitative polymerase chain reaction (qPCR). Additionally, CCK8 experiments and wound healing assays were performed to assess cell viability, migration, and invasion. Finally, online database predictions were used to validate the drug sensitivity of glioma. Results: LINC01842 was more highly expressed in high-grade gliomas, and glioma patients with high expression level of LINC01842 had poorer survival. Additionally, tumor cells with high expression of LINC01842 exhibited stronger tumor characteristics, such as migratory and invasive abilities, as well as tolerance to disulfidptosis. Furthermore, strong binding between LINC01842 and the disulfidptosis-related gene IQGAP1 could be predicted, and their expression levels were positively correlated. Ultimately, drug sensitivity analysis suggested glioma patients with high expression level of LINC01842 were sensitive to eight drugs. Conclusion: High expression of LINC01842 is associated with poor prognosis in glioma patients. By interacting with IQGAP1, the resistance to disulfidptosis of LINC0182 was stronger in glioma, which promotes the progression of glioma. Eight discovered sensitive drugs provided important clues for personalized treatment of glioma.

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The Association of MEG3 lncRNA with Nuclear Speckles in Living Cells

Cited by 6 publications

References 71 publications

Nuclear RNA-related processes modulate the assembly of cytoplasmic RNA granules

Nuclear RNA-related processes modulate the assembly of cytoplasmic RNA granules

CCAT1 lncRNA is chromatin-retained and post-transcriptionally spliced

Long noncoding RNA LINC01842 enhances disulfidptosis resistance and promotes glioma progression via IQGAP1

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