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Mob DNA. 2019 Dec 29;10:51. doi: 10.1186/s13100-019-0196-x. eCollection 2019.

A re-evaluation of LINE-1 ORF2 expression in LNCaP prostate cancer cells.

Author information

1
1Departments of Biochemistry and Molecular Pharmacology, New York University School of Medicine, Alexandria Center for Life Sciences, 450 East 29th Street, Room 323, New York, NY 10016 USA.
2
3Institute of Translational Pharmacology, National Research Council, Rome, Italy.
3
2Departments of Urology, New York University School of Medicine, Alexandria Center for Life Sciences, 450 East 29th Street, Room 323, New York, NY 10016 USA.

Abstract

Background:

We previously examined expression of Long Interspersed Element-1 (LINE-1) in a variety of prostate cancer cells including hormone-dependent LNCaP cells. These studies demonstrated expression and sub-cellular localization of LINE-1 proteins, ORF1p, with antibody 4H1, and ORF2p, with antibody chA1-L1.

Results:

Here we conduct immunoprecipitation/mass spectrometry analysis using chA1-L1 antibody against ORF2p in LNCaP cells. Our results indicate that antigens detected by the antibody include the transcriptional regulator BCLAF1. We show that chA1-L1 recognizes BCLAF1 using siRNA knockdown and overexpression of a tagged BCLAF1. We also show that chA1-L1 antibody recognizes ORF2p in HEK293 cells overexpressing LINE-1. Further, analysis of ORF2p (chA1-L1) and BCLAF1 foci using immunofluorescence in LNCaP cells showed significant colocalization.

Conclusions:

Overall, our findings indicate that chA1-L1 antibody recognizes both BCLAF1 and ORF2p but the majority of antigen recognized in LNCaP cells is BCLAF1.

KEYWORDS:

BCLAF1; LINE-1; ORF2; ORF2 antibody; Prostate cancer

Conflict of interest statement

Competing interestsThe authors declare that they have no competing interests. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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