Micropeptide ASAP encoded by LINC00467 promotes colorectal cancer progression by directly modulating ATP synthase activity
Qiwei Ge, … , Shujie Chen, Liangjing Wang
Qiwei Ge, … , Shujie Chen, Liangjing Wang
Published September 30, 2021
Citation Information: J Clin Invest. 2021;131(22):e152911. https://doi.org/10.1172/JCI152911.
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Research Article Gastroenterology Oncology

Micropeptide ASAP encoded by LINC00467 promotes colorectal cancer progression by directly modulating ATP synthase activity

Abstract

Emerging evidence has shown that open reading frames inside long noncoding RNAs (lncRNAs) could encode micropeptides. However, their roles in cellular energy metabolism and tumor progression remain largely unknown. Here, we identified a 94 amino acid–length micropeptide encoded by lncRNA LINC00467 in colorectal cancer. We also characterized its conservation across higher mammals, localization to mitochondria, and the concerted local functions. This peptide enhanced the ATP synthase construction by interacting with the subunits α and γ (ATP5A and ATP5C), increased ATP synthase activity and mitochondrial oxygen consumption rate, and thereby promoted colorectal cancer cell proliferation. Hence, this micropeptide was termed ATP synthase–associated peptide (ASAP). Furthermore, loss of ASAP suppressed patient-derived xenograft growth with attenuated ATP synthase activity and mitochondrial ATP production. Clinically, high expression of ASAP and LINC00467 predicted poor prognosis of colorectal cancer patients. Taken together, our findings revealed a colorectal cancer–associated micropeptide as a vital player in mitochondrial metabolism and provided a therapeutic target for colorectal cancer.

Authors

Qiwei Ge, Dingjiacheng Jia, Dong Cen, Yadong Qi, Chengyu Shi, Junhong Li, Lingjie Sang, Luo-jia Yang, Jiamin He, Aifu Lin, Shujie Chen, Liangjing Wang

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Figure 1

LINC00467 encodes an uncharacterized peptide, ASAP.

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LINC00467 encodes an uncharacterized peptide, ASAP. (A) A Venn diagram s...
(A) A Venn diagram showing upregulated genes in CRC (GSE22598), ribosomal lncRNAs in HCT116 cells (GSE143263), and translated lncRNA from a previously reported database. (B) The expression of 6 candidate lncRNAs in the TCGA COAD database. (C) Relative levels of 3 candidate lncRNAs in HCT116 cells were determined by polysome profiling followed by RT-qPCR. Data are presented as mean ± SD. n = 3 biologically independent experiments. (D) Data set from GSE22598 indicates LINC00467 expression levels in CRC tissues and adjacent normal tissues (n = 17). Data are presented as a box plot with box and whiskers. Bounds of box show the 25th and 75th percentiles, and the central lines in the box represent the median value. Whiskers show 10th to 90th percentiles. Outlying values, 5.37, 4.38 (N), 8.09, 4.77 (T). Paired samples, 2-sided Student’s t test. **P < 0.01. (E) Five ORF-Flag fusion constructs were transfected into HCT116 cells, and ORF-Flag fusion protein was detected by Western blot analysis. (F) Diagram of Flag fusion constructs. Start codon of ORF1 was mutated to ATT. (G) Indicated constructs were transfected in HCT116 cells, and Flag signal (green) was detected by immunofluorescence. Nuclei were stained with DAPI (blue). Scale bar: 10 μm. (H) Diagram of ORF1 location at the LINC00467 locus and the Flag-tag that was inserted to the end of ORF1. The 6 exons and Poly(A) tail of LINC00467 are shown. (I) ORF-Flag fusion protein (green) was detected by immunofluorescence in HCT116 cells. Nuclei were stained with DAPI (blue). Scale bar: 10 μm. (J) Expression of ASAP peptide (green) in HCT116 cells was detected by immunofluorescence with prepared anti-ASAP antibody. Nuclei were stained with DAPI (blue). Scale bar: 10 μm. Data are representative of 3 independent experiments (E, G, I, and J).