Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023:2607:215-256.
doi: 10.1007/978-1-0716-2883-6_12.

Affinity-Based Interactome Analysis of Endogenous LINE-1 Macromolecules

Luciano H Di Stefano  1 , Leila J Saba  1 , Mehrnoosh Oghbaie  2 , Hua Jiang  2 , Wilson McKerrow  3 , Maria Benitez-Guijarro  4 , Martin S Taylor  5 , John LaCava  6   7
Affiliations

Affinity-Based Interactome Analysis of Endogenous LINE-1 Macromolecules

Luciano H Di Stefano et al. Methods Mol Biol. 2023.

Abstract

During their proliferation and the host's concomitant attempts to suppress it, LINE-1 (L1) retrotransposons give rise to a collection of heterogeneous ribonucleoproteins (RNPs); their protein and RNA compositions remain poorly defined. The constituents of L1-associated macromolecules can differ depending on numerous factors, including, for example, position within the L1 life cycle, whether the macromolecule is productive or under suppression, and the cell type within which the proliferation is occurring. This chapter describes techniques that aid the capture and characterization of protein and RNA components of L1 macromolecules from tissues that natively express them. The protocols described have been applied to embryonal carcinoma cell lines that are popular model systems for L1 molecular biology (e.g., N2102Ep, NTERA-2, and PA-1 cells), as well as colorectal cancer tissues. N2102Ep cells are given as the use case for this chapter; the protocols should be applicable to essentially any tissue exhibiting endogenous L1 expression with minor modifications.

Keywords: Affinity capture; Interactomics; Protein complexes; Retrotransposon; Ribonucleoprotein.

PubMed Disclaimer

References

Bibliography

    1. Ostertag EM, Kazazian HH (2001) Biology of mammalian L1 retrotransposons. Annu Rev Genet 35:501–538 - PubMed
    1. Moran JV, Gilbert N (2002) Mammalian LINE-1 retrotransposons and related elements. In: Craig NLC, Cragie R, Gellert M, Lambowitz AM (eds) Mobile DNA II. American Society of Microbiology, Washington, DC, pp 836–869
    1. Beauregard A, Curcio MJ, Belfort M (2008) The take and give between retrotransposable elements and their hosts. Annu Rev Genet 42:587–617 - PubMed - PMC
    1. Goodier JL, Kazazian HH (2008) Retrotransposons revisited: the restraint and rehabilitation of parasites. Cell 135(1):23–35 - PubMed
    1. Paço A, Adega F, Chaves R (2015) LINE-1 retrotransposons: from “parasite” sequences to functional elements. J Appl Genet 56(1):133–145 - PubMed

Publication types

LinkOut - more resources