Dissection of affinity captured LINE-1 macromolecular complexes

Martin S. Taylor, Ilya Altukhov, Kelly R. Molloy, Paolo Mita, Hua Jiang, Emily M. Adney, Aleksandra Wudzinska, Sana Badri, Dmitry Ischenko, George Eng, Kathleen Burns, David Fenyö, Brian T. Chait, Dmitry Alexeev, Michael P. Rout, Jef D. Boeke, John LaCava

Research output: Contribution to journalArticle

Abstract

Long Interspersed Nuclear Element-1 (LINE-1, L1) is a mobile genetic element active in human genomes. L1-encoded ORF1 and ORF2 proteins bind L1 RNAs, forming ribonucleoproteins (RNPs). These RNPs interact with diverse host proteins, some repressive and others required for the L1 lifecycle. Using differential affinity purifications, quantitative mass spectrometry, and next generation RNA sequencing, we have characterized the proteins and nucleic acids associated with distinctive, enzymatically active L1 macromolecular complexes. Among them, we describe a cytoplasmic intermediate that we hypothesize to be the canonical ORF1p/ORF2p/L1-RNAcontaining RNP, and we describe a nuclear population containing ORF2p, but lacking ORF1p, which likely contains host factors participating in target-primed reverse transcription.

Original languageEnglish (US)
Article numbere30094
JournaleLife
Volume7
DOIs
StatePublished - Jan 8 2018

Fingerprint

Macromolecular Substances
Dissection
Ribonucleoproteins
Long Interspersed Nucleotide Elements
RNA
Interspersed Repetitive Sequences
RNA Sequence Analysis
Proteins
Human Genome
Transcription
Nucleic Acids
Reverse Transcription
Purification
Mass spectrometry
Mass Spectrometry
Genes
Population

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Taylor, M. S., Altukhov, I., Molloy, K. R., Mita, P., Jiang, H., Adney, E. M., ... LaCava, J. (2018). Dissection of affinity captured LINE-1 macromolecular complexes. eLife, 7, [e30094]. https://doi.org/10.7554/eLife.30094

Dissection of affinity captured LINE-1 macromolecular complexes. / Taylor, Martin S.; Altukhov, Ilya; Molloy, Kelly R.; Mita, Paolo; Jiang, Hua; Adney, Emily M.; Wudzinska, Aleksandra; Badri, Sana; Ischenko, Dmitry; Eng, George; Burns, Kathleen; Fenyö, David; Chait, Brian T.; Alexeev, Dmitry; Rout, Michael P.; Boeke, Jef D.; LaCava, John.

In: eLife, Vol. 7, e30094, 08.01.2018.

Research output: Contribution to journalArticle

Taylor, MS, Altukhov, I, Molloy, KR, Mita, P, Jiang, H, Adney, EM, Wudzinska, A, Badri, S, Ischenko, D, Eng, G, Burns, K, Fenyö, D, Chait, BT, Alexeev, D, Rout, MP, Boeke, JD & LaCava, J 2018, 'Dissection of affinity captured LINE-1 macromolecular complexes', eLife, vol. 7, e30094. https://doi.org/10.7554/eLife.30094
Taylor MS, Altukhov I, Molloy KR, Mita P, Jiang H, Adney EM et al. Dissection of affinity captured LINE-1 macromolecular complexes. eLife. 2018 Jan 8;7. e30094. https://doi.org/10.7554/eLife.30094
Taylor, Martin S. ; Altukhov, Ilya ; Molloy, Kelly R. ; Mita, Paolo ; Jiang, Hua ; Adney, Emily M. ; Wudzinska, Aleksandra ; Badri, Sana ; Ischenko, Dmitry ; Eng, George ; Burns, Kathleen ; Fenyö, David ; Chait, Brian T. ; Alexeev, Dmitry ; Rout, Michael P. ; Boeke, Jef D. ; LaCava, John. / Dissection of affinity captured LINE-1 macromolecular complexes. In: eLife. 2018 ; Vol. 7.
@article{19726c49f3014de2acc5137532479bee,
title = "Dissection of affinity captured LINE-1 macromolecular complexes",
abstract = "Long Interspersed Nuclear Element-1 (LINE-1, L1) is a mobile genetic element active in human genomes. L1-encoded ORF1 and ORF2 proteins bind L1 RNAs, forming ribonucleoproteins (RNPs). These RNPs interact with diverse host proteins, some repressive and others required for the L1 lifecycle. Using differential affinity purifications, quantitative mass spectrometry, and next generation RNA sequencing, we have characterized the proteins and nucleic acids associated with distinctive, enzymatically active L1 macromolecular complexes. Among them, we describe a cytoplasmic intermediate that we hypothesize to be the canonical ORF1p/ORF2p/L1-RNAcontaining RNP, and we describe a nuclear population containing ORF2p, but lacking ORF1p, which likely contains host factors participating in target-primed reverse transcription.",
author = "Taylor, {Martin S.} and Ilya Altukhov and Molloy, {Kelly R.} and Paolo Mita and Hua Jiang and Adney, {Emily M.} and Aleksandra Wudzinska and Sana Badri and Dmitry Ischenko and George Eng and Kathleen Burns and David Feny{\"o} and Chait, {Brian T.} and Dmitry Alexeev and Rout, {Michael P.} and Boeke, {Jef D.} and John LaCava",
year = "2018",
month = "1",
day = "8",
doi = "10.7554/eLife.30094",
language = "English (US)",
volume = "7",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",

}

TY - JOUR

T1 - Dissection of affinity captured LINE-1 macromolecular complexes

AU - Taylor, Martin S.

AU - Altukhov, Ilya

AU - Molloy, Kelly R.

AU - Mita, Paolo

AU - Jiang, Hua

AU - Adney, Emily M.

AU - Wudzinska, Aleksandra

AU - Badri, Sana

AU - Ischenko, Dmitry

AU - Eng, George

AU - Burns, Kathleen

AU - Fenyö, David

AU - Chait, Brian T.

AU - Alexeev, Dmitry

AU - Rout, Michael P.

AU - Boeke, Jef D.

AU - LaCava, John

PY - 2018/1/8

Y1 - 2018/1/8

N2 - Long Interspersed Nuclear Element-1 (LINE-1, L1) is a mobile genetic element active in human genomes. L1-encoded ORF1 and ORF2 proteins bind L1 RNAs, forming ribonucleoproteins (RNPs). These RNPs interact with diverse host proteins, some repressive and others required for the L1 lifecycle. Using differential affinity purifications, quantitative mass spectrometry, and next generation RNA sequencing, we have characterized the proteins and nucleic acids associated with distinctive, enzymatically active L1 macromolecular complexes. Among them, we describe a cytoplasmic intermediate that we hypothesize to be the canonical ORF1p/ORF2p/L1-RNAcontaining RNP, and we describe a nuclear population containing ORF2p, but lacking ORF1p, which likely contains host factors participating in target-primed reverse transcription.

AB - Long Interspersed Nuclear Element-1 (LINE-1, L1) is a mobile genetic element active in human genomes. L1-encoded ORF1 and ORF2 proteins bind L1 RNAs, forming ribonucleoproteins (RNPs). These RNPs interact with diverse host proteins, some repressive and others required for the L1 lifecycle. Using differential affinity purifications, quantitative mass spectrometry, and next generation RNA sequencing, we have characterized the proteins and nucleic acids associated with distinctive, enzymatically active L1 macromolecular complexes. Among them, we describe a cytoplasmic intermediate that we hypothesize to be the canonical ORF1p/ORF2p/L1-RNAcontaining RNP, and we describe a nuclear population containing ORF2p, but lacking ORF1p, which likely contains host factors participating in target-primed reverse transcription.

UR - http://www.scopus.com/inward/record.url?scp=85043495559&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85043495559&partnerID=8YFLogxK

U2 - 10.7554/eLife.30094

DO - 10.7554/eLife.30094

M3 - Article

VL - 7

JO - eLife

JF - eLife

SN - 2050-084X

M1 - e30094

ER -