A potential role for the nucleous in L1 retrotransposition

John L. Goodier; Eric M. Ostertag; Kurt A. Engleka; Maria C. Seleme; Haig H. Kazazian

doi:10.1093/hmg/ddh118

A potential role for the nucleous in L1 retrotransposition

John L. Goodier, Eric M. Ostertag, Kurt A. Engleka, Maria C. Seleme, Haig H. Kazazian

Research output: Contribution to journal › Article › peer-review

85 Scopus citations

Abstract

Determining the subcellular localization of the L1 ORF2 protein (ORF2p) has been impossible to date because of technical limitations in detecting either endogenous or overexpressed forms of the protein. Here we report visualization of the full-length ORF2p in cultured human cells following expression in a modified vaccinia virus/T7 RNA polymerase (MVA/T7RP) system. The MVA/T7RP system was used to ascertain subcellular localization of L1 ORF1p and ORF2p both as fusions with green fluorescent protein and by immunocytochemistry. Full-length ORF2p was predominantly cytoplasmic, while carboxy-terminal-deleted ORF2p localized additionally to the nucleolus. We mapped a functional nucleolar localization signal in ORF2p. ORF1p appeared in the cytoplasm with a speckled pattern and colocalized with ORF2p in nucleoli in a subset of cells. These findings help explain the presence of chimeras between L1s and small RNA gene sequences recently discovered in the human genome.

Original language	English (US)
Pages (from-to)	1041-1048
Number of pages	8
Journal	Human molecular genetics
Volume	13
Issue number	10
DOIs	https://doi.org/10.1093/hmg/ddh118
State	Published - May 15 2004
Externally published	Yes

ASJC Scopus subject areas

Molecular Biology
Genetics
Genetics(clinical)

Access to Document

10.1093/hmg/ddh118

Cite this

@article{5a5fd3be65e64cab9f7ae886975e9197,

title = "A potential role for the nucleous in L1 retrotransposition",

abstract = "Determining the subcellular localization of the L1 ORF2 protein (ORF2p) has been impossible to date because of technical limitations in detecting either endogenous or overexpressed forms of the protein. Here we report visualization of the full-length ORF2p in cultured human cells following expression in a modified vaccinia virus/T7 RNA polymerase (MVA/T7RP) system. The MVA/T7RP system was used to ascertain subcellular localization of L1 ORF1p and ORF2p both as fusions with green fluorescent protein and by immunocytochemistry. Full-length ORF2p was predominantly cytoplasmic, while carboxy-terminal-deleted ORF2p localized additionally to the nucleolus. We mapped a functional nucleolar localization signal in ORF2p. ORF1p appeared in the cytoplasm with a speckled pattern and colocalized with ORF2p in nucleoli in a subset of cells. These findings help explain the presence of chimeras between L1s and small RNA gene sequences recently discovered in the human genome.",

author = "Goodier, {John L.} and Ostertag, {Eric M.} and Engleka, {Kurt A.} and Seleme, {Maria C.} and Kazazian, {Haig H.}",

note = "Funding Information: The authors thank L.S. Wyatt and B. Moss (NIH) for MVA/ T7RP virus and A. Farley for technical assistance. This work was supported in part by NIH Grant 5R21GM065522-02 to J.L.G. E.M.O was supported by a HHMI predoctoral fellowship.",

year = "2004",

month = may,

day = "15",

doi = "10.1093/hmg/ddh118",

language = "English (US)",

volume = "13",

pages = "1041--1048",

journal = "Human molecular genetics",

issn = "0964-6906",

publisher = "Oxford University Press",

number = "10",

}

TY - JOUR

T1 - A potential role for the nucleous in L1 retrotransposition

AU - Goodier, John L.

AU - Ostertag, Eric M.

AU - Engleka, Kurt A.

AU - Seleme, Maria C.

AU - Kazazian, Haig H.

N1 - Funding Information: The authors thank L.S. Wyatt and B. Moss (NIH) for MVA/ T7RP virus and A. Farley for technical assistance. This work was supported in part by NIH Grant 5R21GM065522-02 to J.L.G. E.M.O was supported by a HHMI predoctoral fellowship.

PY - 2004/5/15

Y1 - 2004/5/15

N2 - Determining the subcellular localization of the L1 ORF2 protein (ORF2p) has been impossible to date because of technical limitations in detecting either endogenous or overexpressed forms of the protein. Here we report visualization of the full-length ORF2p in cultured human cells following expression in a modified vaccinia virus/T7 RNA polymerase (MVA/T7RP) system. The MVA/T7RP system was used to ascertain subcellular localization of L1 ORF1p and ORF2p both as fusions with green fluorescent protein and by immunocytochemistry. Full-length ORF2p was predominantly cytoplasmic, while carboxy-terminal-deleted ORF2p localized additionally to the nucleolus. We mapped a functional nucleolar localization signal in ORF2p. ORF1p appeared in the cytoplasm with a speckled pattern and colocalized with ORF2p in nucleoli in a subset of cells. These findings help explain the presence of chimeras between L1s and small RNA gene sequences recently discovered in the human genome.

AB - Determining the subcellular localization of the L1 ORF2 protein (ORF2p) has been impossible to date because of technical limitations in detecting either endogenous or overexpressed forms of the protein. Here we report visualization of the full-length ORF2p in cultured human cells following expression in a modified vaccinia virus/T7 RNA polymerase (MVA/T7RP) system. The MVA/T7RP system was used to ascertain subcellular localization of L1 ORF1p and ORF2p both as fusions with green fluorescent protein and by immunocytochemistry. Full-length ORF2p was predominantly cytoplasmic, while carboxy-terminal-deleted ORF2p localized additionally to the nucleolus. We mapped a functional nucleolar localization signal in ORF2p. ORF1p appeared in the cytoplasm with a speckled pattern and colocalized with ORF2p in nucleoli in a subset of cells. These findings help explain the presence of chimeras between L1s and small RNA gene sequences recently discovered in the human genome.

UR - http://www.scopus.com/inward/record.url?scp=2542580854&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2542580854&partnerID=8YFLogxK

U2 - 10.1093/hmg/ddh118

DO - 10.1093/hmg/ddh118

M3 - Article

C2 - 15028673

AN - SCOPUS:2542580854

SN - 0964-6906

VL - 13

SP - 1041

EP - 1048

JO - Human molecular genetics

JF - Human molecular genetics

IS - 10

ER -

A potential role for the nucleous in L1 retrotransposition

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this