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不同HeLa细胞系中反转录转座的可变模式为SINE RNA移动过程提供了机制上的见解。

Variable patterns of retrotransposition in different HeLa strains provide mechanistic insights into SINE RNA mobilization processes.

作者信息

Moldovan John B, Kopera Huira C, Liu Ying, Garcia-Canadas Marta, Catalina Purificacion, Leone Paola E, Sanchez Laura, Kitzman Jacob O, Kidd Jeffrey M, Garcia-Perez Jose Luis, Moran John V

出版信息

bioRxiv. 2024 May 3:2024.05.03.592410. doi: 10.1101/2024.05.03.592410.

DOI:10.1101/2024.05.03.592410
PMID:38746229
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11092746/
Abstract

elements are non-autonomous Short INterspersed Elements (SINEs) derived from the gene that are present at over one million copies in human genomic DNA. mobilizes by a mechanism known as retrotransposition, which requires the Long INterspersed Element-1 (LINE-1 or L1) -encoded protein (ORF2p). Here, we demonstrate that HeLa strains differ in their capacity to support retrotransposition. Human elements retrotranspose efficiently in HeLa-HA and HeLa-CCL2 ( -permissive) strains, but not in HeLa-JVM or HeLa-H1 ( -nonpermissive) strains. A similar pattern of retrotransposition was observed for other -derived SINEs and -derived SINEs. In contrast, mammalian LINE-1s, a zebrafish LINE, a human - ( ) element, and an -containing messenger RNA can retrotranspose in all four HeLa strains. Using an reverse transcriptase-based assay, we show that RNAs associate with ORF2p and are converted into cDNAs in both -permissive and -nonpermissive HeLa strains, suggesting that - and -derived SINE RNAs use strategies to 'hijack' L1 ORF2p that are distinct from those used by elements and -containing mRNAs. These data further suggest ORF2p associates with the RNA poly(A) tract in both -permissive and -nonpermissive HeLa strains, but that retrotransposition is blocked after this critical step in -nonpermissive HeLa strains.

摘要

元件是非自主的短散在元件(SINEs),源自该基因,在人类基因组DNA中存在超过一百万个拷贝。通过一种称为逆转录转座的机制进行移动,这需要长散在元件-1(LINE-1或L1)编码的蛋白质(ORF2p)。在这里,我们证明HeLa细胞系在支持逆转录转座的能力上存在差异。人类元件在HeLa-HA和HeLa-CCL2(允许)细胞系中能高效逆转录转座,但在HeLa-JVM或HeLa-H1(非允许)细胞系中则不能。对于其他源自的SINEs和源自的SINEs,也观察到了类似的逆转录转座模式。相比之下,哺乳动物的LINE-1s、一种斑马鱼的LINE、人类的 - ( )元件以及一种含的信使RNA在所有四种HeLa细胞系中都能逆转录转座。使用基于逆转录酶的检测方法,我们表明RNA与ORF2p结合,并在允许和非允许的HeLa细胞系中都被转化为cDNA,这表明源自和源自的SINE RNA使用了与元件和含mRNA不同的策略来“劫持”L1 ORF2p。这些数据进一步表明,在允许和非允许的HeLa细胞系中,ORF2p都与RNA的聚腺苷酸尾结合,但在非允许的HeLa细胞系中,在这一关键步骤之后逆转录转座被阻断。

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Variable patterns of retrotransposition in different HeLa strains provide mechanistic insights into SINE RNA mobilization processes.不同HeLa细胞系中反转录转座的可变模式为SINE RNA移动过程提供了机制上的见解。
bioRxiv. 2024 May 3:2024.05.03.592410. doi: 10.1101/2024.05.03.592410.
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