Abstract
The p53-induced long noncoding RNA (lncRNA) lincRNA-p21 is proposed to act in cis to promote p53-dependent expression of the neighboring cell cycle gene, Cdkn1a/p21. The molecular mechanism through which the transcribed lincRNA-p21 regulatory locus activates p21 expression remains poorly understood. To elucidate the functional elements of cis-regulation, we generate a series of genetic models that disrupt DNA regulatory elements, the transcription of lincRNA-p21, or the accumulation of mature lincRNA-p21. Unexpectedly, we determine that full-length transcription, splicing, and accumulation of lincRNA-p21 are dispensable for the chromatin organization of the locus and for cis-regulation. Instead, we find that production of lincRNA-p21 through conserved regions in exon 1 of lincRNA-p21 promotes cis-activation. These findings demonstrate that the activation of nascent transcription from this lncRNA locus, but not the generation or accumulation of a mature lncRNA transcript, is necessary to enact local gene expression control.
| Original language | English (US) |
|---|---|
| Article number | 110687 |
| Journal | Cell reports |
| Volume | 39 |
| Issue number | 3 |
| DOIs | |
| State | Published - Apr 19 2022 |
Funding
We thank Elena Martinez, Emily Dangelmaier, and Zara Malik for helpful comments. We are grateful to Christiane Olivero for TWI design and Rick Maser from Jackson Laboratories for lincRNA-p21 mutant founders generation. This work was supported in part by LCRF (N.D.), IRG-ACS 58-012-58 (N.D.), The V Foundation (N.D.), the Pew-Stewart Foundation (N.D.), NIH R37CA230580 grant (N.D.), NIH R01GM137117 (M.D.S.), and by an SPORE in Lung Cancer (Yale University, NIH P50CA196530). L.W. was supported by NIH T32GM007223. L.W. and N.D. designed the study, performed experiments, analyzed data, generated figures, and prepared the manuscript. M.J. performed experiments and analyzed data. A.W. enabled the generation of the mouse models with input from L.W. and N.D. J.Z. and M.S. performed analysis of metabolically labeled RNA. All authors approved the final manuscript. The authors declare no competing interests. We thank Elena Martinez, Emily Dangelmaier, and Zara Malik for helpful comments. We are grateful to Christiane Olivero for TWI design and Rick Maser from Jackson Laboratories for lincRNA-p21 mutant founders generation. This work was supported in part by LCRF (N.D.), IRG-ACS 58-012-58 (N.D.), The V Foundation (N.D.), the Pew-Stewart Foundation (N.D.), NIH R37CA230580 grant (N.D.), NIH R01GM137117 (M.D.S.), and by an SPORE in Lung Cancer (Yale University, NIH P50CA196530 ). L.W. was supported by NIH T32GM007223 .
Keywords
- CP: Molecular biology
- cis-regulation
- genetic models
- lincRNA-p21
- long noncoding RNA
- transcription
ASJC Scopus subject areas
- General Biochemistry, Genetics and Molecular Biology