A Marcello

A Sabo

Chiara Vardabasso

Lara Manganaro

Maria Ines Gutierrez

Marina Lusic

Mauro Giacca

S Briers

Retrovirology

English

PubMed

Springer - Publisher Connector

BioMed Central
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Retrovirology
Open AccessPoster presentation
Spatial juxtaposition of HIV-1 provirus with PML and KAKA bodies 
as revealed by 3D Immuno DNA FISH
Marina Lusic*, Lara Manganaro, Chiara Vardabasso, Maria Ines Gutierrez and 
Mauro Giacca
Address: Molecular Medicine Laboratory, ICGEB, Trieste, Italy
* Corresponding author    
Various members of tripartite motif (TRIM) protein fam-
ily display antiviral properties, targeting retroviruses in
particular. The potential activity of TRIM19, better known
as promyelocytic leukemia protein (PML) against several
viruses has been well documented, yet it's role in HIV-1
infection remains elusive.
One of the most important cellular partners of HIV-1, P-
TEFb kinase complex, composed of CDK9 and CyclinT1
plays a crucial role in regulation of HIV-1 transcription.
We have previously demonstrated that both members of
P-TEFb interact with PML protein and localize inside the
PML Nuclear Bodies (PML NB) [1,2]. In particular, we
found that the acetylated and enzymaticaly inactive form
of CDK9 binds PML and can be separated with PML in the
insoluble chromatin fraction. Our ChIP analysis revealed
that acetylated form of CDK9 localizes to the integrated
and transcriptionally silent viral genome, thus indicating
that PML might have yet unidentified role in regulation of
HIV-1 infection and latency [2].
We indeed identified PML protein bound to the silent
viral genome in Jurkat cells harboring single integrations
of HIV-1 provirus. The distribution of PML protein along
the viral genome could be correlated with bimethylated
lysine 9 (K9) on Histone 3 (2MetK9 H3), a mark of facul-
tative heterochromatin. Transcriptional activation of the
provirus with TNF-α led to a displacement of PML protein
from the genome and to a substantial loss of 2MetK9H3
mark. By three-dimensional immuno fluorescent in situ
hybridization (3D Immuno DNA FISH) that we devel-
oped we visualized the viral genome in close proximity to
PML NB and we characterized this position as adjacent.
Within the population of cells tested in the silenced state,
40% of the cells showed proximity at the distance of less
that 0.6μm. PML NB were recently described to have a
close spatial relationship with the so called KAKA foci,
characterized by the presence of KAP-1 (or Trim28) and
KRAB-Zinc Finger proteins [3] thus prompting us to ana-
lyse the relationship of the silent HIV-1 provirus with
these nuclear structures. We detected numerous PML and
KAKA foci in both Jurkat cells (J-LAT model of latency)
and primary human CD4+T lymphocytes, with HIV-1
residing either in close proximity to PML or to KAKA foci
(~40% showing less than 0.6μm distance) in J-LAT latency
model. Whether this spatial localization of HIV-1 provirus
is correlated to silencing of the virus genome or is a gen-
eral feature of HIV-1 integration and replication remains
still to be determined.
References
1. Marcello A, et al.: Recruitment of human cyclin T1 to nuclear
bodies through direct interaction with the PML protein.
Embo J 2003, 22(9):2156-66.
2. Sabo A, et al.: Acetylation of conserved lysines in the catalytic
core of cyclin-dependent kinase 9 inhibits kinase activity and
regulates transcription.  Mol Cell Biol 2008, 28(7):2201-12.
3. Briers S, et al.: KRAB zinc-finger proteins localise to novel
KAP1-containing foci that are adjacent to PML nuclear bod-
ies.  J Cell Sci 2009, 122(Pt 7):937-46.
from Frontiers of Retrovirology: Complex retroviruses, retroelements and their hosts
Montpellier, France. 21-23 September 2009
Published: 24 September 2009
Retrovirology 2009, 6(Suppl 2):P52 doi:10.1186/1742-4690-6-S2-P52
<supplement> <title> <p>Frontiers of Retrovirology: Complex retroviruses, retroelements and their hosts</p> </title> <note>Meeting abstracts - A single PDF containing all abstracts in this Supplement is available <a href="http://www.biomedcentral.com/content/files/pdf/1742-4690-6-S2-full.pdf">here</a>.</note> <url>http://www.biomedcentral.com/content/pdf/1742-4690-6-S2-info.pdf</url> </supplement>
This abstract is available from: http://www.retrovirology.com/content/6/S2/P52
© 2009 Lusic et al; licensee BioMed Central Ltd. 


Spatial juxtaposition of HIV-1 provirus with PML and KAKA bodies as revealed by 3D Immuno DNA FISH

Crossref

Acetylation of conserved lysines in the catalytic core of cyclin-dependent kinase 9 inhibits kinase activity and regulates transcription. Mol Cell Biol

et al.: Recruitment of human cyclin T1 to nuclear bodies through direct interaction with the PML protein. Embo J

KRAB zinc-finger proteins localise to novel KAP1-containing foci that are adjacent to PML nuclear bodies. J Cell Sci 2009, 122(Pt 7):937-46. from Frontiers of Retrovirology: Complex retroviruses, retroelements and their hosts

http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2767037

Spatial juxtaposition of HIV-1 provirus with PML and KAKA bodies as revealed by 3D Immuno DNA FISH

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