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Differential Interaction of HIV-1 Integrase and JPO2 with the C Terminus of LEDGF/p75
- Source :
- Journal of Molecular Biology. 372:407-421
- Publication Year :
- 2007
- Publisher :
- Elsevier BV, 2007.
-
Abstract
- The transcriptional co-activator lens epithelium-derived growth factor (LEDGF) has been shown to protect cells against environmental stress. The protein has been implicated in auto-immunity and cancer, and is present in cells as the p52 or p75 splice variant. Recently, LEDGF/p75, but not p52, was identified as the prominent interaction partner of human immunodeficiency virus type 1 (HIV-1) integrase. This interaction of HIV-1 integrase with the C-terminal integrase-binding domain of LEDGF/p75 is crucial for HIV-1 replication. To gain insight into the cell biology of LEDGF/p75, we were interested in identifying cellular binding partners of its C-terminal domain. By yeast-two-hybrid screening with a CEMC7 cDNA-library, we were able to identify JPO2 as a binding partner of the C-terminal part of LEDGF/p75. The specific interaction between JPO2 and LEDGF/p75 was verified by pull-down, AlphaScreen, and co-immunoprecipitation. Competition assays using recombinant proteins show a mutually exclusive binding of either JPO2 or HIV-1 integrase to LEDGF/p75. However, differing mechanisms of binding were suggested by continuing interaction of JPO2 with some LEDGF/p75 mutants (I365A, D366A, F406A) that are totally defective for interaction with HIV-1 integrase. This finding is of significance for the development of specific inhibitors targeting only the interaction between LEDGF/p75 and HIV-1 integrase, without disturbing interaction with other cellular factors. Over-expression of JPO2 resulted in a modest but reproducible inhibition of HIV-1 replication, consistent with competition between integrase and JPO2 for binding to LEDGF/p75. Furthermore, JPO2 over-expression activated transcription from the HIV-1 LTR.
- Subjects :
- musculoskeletal diseases
medicine.medical_treatment
Molecular Sequence Data
Mutant
Gene Expression
HIV Integrase
Biology
Virus Replication
Binding, Competitive
law.invention
Structural Biology
Transcription (biology)
law
medicine
Animals
Humans
Amino Acid Sequence
Molecular Biology
Genetics
C-terminus
Growth factor
Alternative splicing
Integrase
Repressor Proteins
Zinc
Multiprotein Complexes
HIV-1
biology.protein
Recombinant DNA
Hiv 1 integrase
Intercellular Signaling Peptides and Proteins
HeLa Cells
Protein Binding
Subjects
Details
- ISSN :
- 00222836
- Volume :
- 372
- Database :
- OpenAIRE
- Journal :
- Journal of Molecular Biology
- Accession number :
- edsair.doi.dedup.....0b42db6d3438e6b48d92b2c4a7031857