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Bacteriophage DNA glucosylation impairs target DNA binding by type I and II but not by type V CRISPR–Cas effector complexes
Vlot, Marnix ; Houkes, Joep ; Lochs, Silke J.A. ; Swarts, Daan C. ; Zheng, Peiyuan ; Kunne, Tim ; Mohanraju, Prarthana ; Anders, Carolin ; Jinek, Martin ; Oost, John Van Der; Dickman, Mark J. ; Brouns, Stan J.J. - \ 2018
Nucleic acids research 46 (2018)2. - ISSN 0305-1048 - p. 873 - 885.
Prokaryotes encode various host defense systems that provide protection against mobile genetic elements. Restriction–modification (R–M) and CRISPR–Cas systems mediate host defense by sequence specific targeting of invasive DNA. T-even bacteriophages employ covalent modifications of nucleobases to avoid binding and therefore cleavage of their DNA by restriction endonucleases. Here, we describe that DNA glucosylation of bacteriophage genomes affects interference of some but not all CRISPR–Cas systems. We show that glucosyl modification of 5-hydroxymethylated cytosines in the DNA of bacteriophage T4 interferes with type I-E and type II-A CRISPR–Cas systems by lowering the affinity of the Cascade and Cas9–crRNA complexes for their target DNA. On the contrary, the type V-A nuclease Cas12a (also known as Cpf1) is not impaired in binding and cleavage of glucosylated target DNA, likely due to a more open structural architecture of the protein. Our results suggest that CRISPR–Cas systems have contributed to the selective pressure on phages to develop more generic solutions to escape sequence specific host defense systems.
Alternative splicing of ALCAM enables tunable regulation of cell-cell adhesion through differential proteolysis
Hebron, Katie E. ; Li, Elizabeth Y. ; Arnold Egloff, Shanna A. ; Lersner, Ariana K. Von; Taylor, Chase ; Houkes, Joep ; Flaherty, David K. ; Eskaros, Adel ; Stricker, Thomas P. ; Zijlstra, Andries - \ 2018
Scientific Reports 8 (2018)1. - ISSN 2045-2322
While many adhesion receptors are known to influence tumor progression, the mechanisms by which they dynamically regulate cell-cell adhesion remain elusive. We previously identified Activated Leukocyte Cell Adhesion Molecule (ALCAM) as a clinically relevant driver of metastasis and hypothesized that a tunable mechanism of ectodomain shedding regulates its contribution to dissemination. To test this hypothesis, we examined an under-explored ALCAM splice variant (ALCAM-Iso2) and demonstrated that loss of the membrane-proximal region of ALCAM (exon 13) increased metastasis four-fold. Mechanistic studies identified a novel MMP14-dependent membrane distal cleavage site in ALCAM-Iso2, which mediated a ten-fold increase in shedding, thereby decreasing cellular cohesion. Importantly, the loss of cohesion is not limited to the cell capable of shedding because the released extracellular domain diminished cohesion of non-shedding cells through disruption of ALCAM-ALCAM interactions. ALCAM-Iso2-dominated expression in bladder cancer tissue, compared to normal bladder, further emphasizes that ALCAM alternative splicing may contribute to clinical disease progression. The requirement for both the loss of exon 13 and the gain of metalloprotease activity suggests that ALCAM shedding and concomitant regulation of tumor cell adhesion is a locally tunable process.
Micromorfologie
Kooistra, M.J. - \ 2008
In: Ypenburg-locatie 4, een nederzetting met grafveld uit het Midden-Neolithicum in het West-Nederlandse Kustgebied / Koot, J.M., Bruning, L., Houkes, R.A., Leiden : Hazenberg Archeologie - ISBN 9789080853454 - p. 59 - 78.
Visualization and quantification of compounds in plant material using spectral image analysis
Polder, G. ; Heijden, G.W.A.M. van der; Young, I.T. - \ 1999
In: Abstracts of the 2nd ASCI Imaging workshop, Enschede, The Netherlands, 29-31 March 1999. - Enschede : ASCI - p. 27 - 28.
A surface Relief Meter Based on Triocular Vision
Sablik, P.W. ; Balendonck, J. ; Ernst, V.G.S. ; Houkes, Z. ; Regtien, P.P.L. - \ 1995
In: Fifth International Conference on Image processing and its Applications, Edinburgh, UK, 2-6 July 1995, pp. 857 - p. 281 - 285.
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