T.B. and G.S. contributed equally to this work.
An L2 SUMO interacting motif is important for PML localization and infection of human papillomavirus type 16
Version of Record online: 12 MAR 2014
© 2014 John Wiley & Sons Ltd
Volume 16, Issue 8, pages 1179–1200, August 2014
How to Cite
Bund, T., Spoden, G. A., Koynov, K., Hellmann, N., Boukhallouk, F., Arnold, P., Hinderberger, D. and Florin, L. (2014), An L2 SUMO interacting motif is important for PML localization and infection of human papillomavirus type 16. Cellular Microbiology, 16: 1179–1200. doi: 10.1111/cmi.12271
- Issue online: 26 JUL 2014
- Version of Record online: 12 MAR 2014
- Accepted manuscript online: 21 JAN 2014 04:33AM EST
- Manuscript Accepted: 14 JAN 2014
- Manuscript Revised: 8 JAN 2014
- Manuscript Received: 3 JUN 2013
- Max Planck Graduate Center
- Johannes Gutenberg University Mainz
- University Mainz
- German Research Foundation. Grant Number: SFB490/D2
Fig. S1. Sequence alignment of L2 in different high-risk HPVs. The potential SIMs at positions 235–238, 408–411 and 427–431 show no or only moderate sequence conservation of the hydrophobic core region. The potential double SIMs at positions 105–109, 145–148, 284–289 share a very high conservation. Especially high sequence conservation was found for the L2 region around the potential double SIM at position 284–289 including the most promising IVAL hydrophobic core motif and, flanking the SIM, conserved negatively charged amino acids (red) as well as a potential casein kinase 2 (CK2) phosphorylation site at position 295/6 (TS). Colour code as in A. Sequence alignment performed with ClustalW2 (http://www.ebi.ac.uk) (Larkin et al., 2007; Goujon et al., 2010), edited with Jalview (http://www.jalview.org) (Waterhouse et al., 2009).
Fig. S2. L2 ΔSCM interacts with GFP-SUMO2 in immunoprecipitation. After coexpression of wt L2 or L2 ΔSCM with GFP-SUMO2 and following immunoprecipitation of GFP-SUMO2, coprecipitation of both L2 wt and L2 ΔSCM was observed.
Fig. S3. L2 colocalization with PML-NBs and GFP-SUMO1/2.
A. Overexpressed L2 D280/2/5N showed a wild-type colocalization with PML-NBs and overexpressed GFP-SUMO2. Colocalization levels were slightly reduced for L2 295/6A.
B. After coexpression of L2 wt and L2 ΔSCM there was a clear colocalization of L2 with PML and coexpressed GFP-SUMO1, which was not observed with L2 286-9A.
Fig. S4. L2 antibody-induced dissociation of L2 and SL-SUMO1/2. EPR samples containing equimolar amounts of L2 and SL-SUMO1 (A) or SL-SUMO2 (B) were incubated with a highly specific L2 antibody (L2-1). The CW EPR spectra obtained after L2 antibody addition featured significantly decreased 2Apar values (52.8–54.5 G, see dashed lines) when compared with 2Apar values from spectra taken before antibody addition (56.9–60 G).
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