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LEDGF/p75 is essential for nuclear and chromosomal targeting of HIV-1 integrase in human cells
- Source :
- The Journal of biological chemistry. 278(35)
- Publication Year :
- 2003
-
Abstract
- We have reported that human immunodeficiency virus type 1 (HIV-1) integrase (IN) forms a specific nuclear complex with human lens epithelium-derived growth factor/transcription co-activator p75 (LEDGF/p75) protein. We now studied the IN-LEDGF/p75 interaction and nuclear import of IN in living cells using fusions of IN and LEDGF/p75 with enhanced green fluorescent protein and far-red fluorescent protein HcRed1. We show that both the N-terminal zinc binding domain and the central core domains of IN are involved in the interaction with LEDGF/p75. Both domains are essential for nuclear localization of IN as well as for the association of IN with condensed chromosomes during mitosis. However, upon overexpression of LEDGF/p75, the core domain fragment of IN was recruited to the nuclei and mitotic chromosomes with a distribution pattern characteristic of the full-length protein, indicating that it harbors the main determinant for interaction with LEDGF/p75. Although the C-terminal domain of IN was dispensable for nuclear/chromosomal localization, a fusion of the C-terminal IN fragment with enhanced green fluorescent protein was found exclusively in the nucleus, with a diffuse nuclear/nucleolar distribution, suggesting that the C-terminal domain may also play a role in the nuclear import of IN. In contrast to LEDGF/p75, its alternative splice variant, p52, did not interact with HIV-1 IN in vitro and in living cells. Finally, RNA interference-mediated knock-down of endogenous LEDGF/p75 expression abolished nuclear/chromosomal localization of IN. We conclude, therefore, that the interaction with LEDGF/p75 accounts for the karyophilic properties and chromosomal targeting of HIV-1 IN.
- Subjects :
- Cytoplasm
Recombinant Fusion Proteins
HIV integration
Blotting, Western
Molecular Sequence Data
Active Transport, Cell Nucleus
Mitosis
HIV Integrase
Biology
Transfection
Biochemistry
PSIP1
Transcription (biology)
medicine
Humans
RNA, Small Interfering
Fluorescent Antibody Technique, Indirect
Molecular Biology
Cells, Cultured
Cell Nucleus
Microscopy, Confocal
Base Sequence
Models, Genetic
Cell Biology
Molecular biology
Integrase
Protein Structure, Tertiary
Cell nucleus
Alternative Splicing
Zinc
medicine.anatomical_structure
Microscopy, Fluorescence
Mutation
biology.protein
Intercellular Signaling Peptides and Proteins
Nuclear transport
Nuclear localization sequence
HeLa Cells
Plasmids
Protein Binding
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 278
- Issue :
- 35
- Database :
- OpenAIRE
- Journal :
- The Journal of biological chemistry
- Accession number :
- edsair.doi.dedup.....95aad9197939fbf768d9e09092392100