{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T11:55:59Z","timestamp":1790855759440,"version":"4.1.0"},"reference-count":31,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5582","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[2002,8,2]]},"abstract":"<jats:p>The passage of large-sized herpesviral capsids through the nuclear lamina and the inner nuclear membrane to leave the nucleus requires a dissolution of the nuclear lamina. Here, we report on the functions of M50\/p35, a \u03b2-herpesviral protein of murine cytomegalovirus. M50\/p35 inserts into the inner nuclear membrane and is aggregated by a second viral protein, M53\/p38, to form the capsid docking site. M50\/p35 recruits the cellular protein kinase C for phosphorylation and dissolution of the nuclear lamina, suggesting that herpesviruses target a critical element of nuclear architecture.<\/jats:p>","DOI":"10.1126\/science.1071506","type":"journal-article","created":{"date-parts":[[2002,10,1]],"date-time":"2002-10-01T15:34:46Z","timestamp":1033486486000},"page":"854-857","source":"Crossref","is-referenced-by-count":239,"title":["Cytomegalovirus Recruitment of Cellular Kinases to Dissolve the Nuclear Lamina"],"prefix":"10.1126","volume":"297","author":[{"given":"Walter","family":"Muranyi","sequence":"first","affiliation":[{"name":"Genzentrum and Max-von-Pettenkofer Institut, Ludwig-Maximilians-Universita\u0308t Mu\u0308nchen, 80336 Mu\u0308nchen, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ju\u0308rgen","family":"Haas","sequence":"additional","affiliation":[{"name":"Genzentrum and Max-von-Pettenkofer Institut, Ludwig-Maximilians-Universita\u0308t Mu\u0308nchen, 80336 Mu\u0308nchen, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Markus","family":"Wagner","sequence":"additional","affiliation":[{"name":"Genzentrum and Max-von-Pettenkofer Institut, Ludwig-Maximilians-Universita\u0308t Mu\u0308nchen, 80336 Mu\u0308nchen, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georg","family":"Krohne","sequence":"additional","affiliation":[{"name":"Division of Electron Microscopy, Biocenter of the University of Wu\u0308rzburg, 97074 Wu\u0308rzburg, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulrich H.","family":"Koszinowski","sequence":"additional","affiliation":[{"name":"Genzentrum and Max-von-Pettenkofer Institut, Ludwig-Maximilians-Universita\u0308t Mu\u0308nchen, 80336 Mu\u0308nchen, Germany."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-5699(00)01699-6"},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/378636a0"},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.76.4.1537-1547.2002"},{"key":"e_1_3_1_5_2","unstructured":"E. S. Mocarski Jr. in Fields Virology B. N. Fields D. M. Knipe P. M. Howley Eds. (Lippincott-Raven Philadelphia ed. 3 1996) pp. 2447\u20132492."},{"key":"e_1_3_1_6_2","first-page":"299","volume":"54","author":"Radsak K. D.","year":"1991","unstructured":"Radsak K. D., Brucher K. H., Georgatos S. D., Eur. J. Cell Biol. 54, 299 (1991).","journal-title":"Eur. J. Cell Biol."},{"key":"e_1_3_1_7_2","unstructured":"W. Muranyi J. Haas M. Wagner G. Krohne U. H. Koszinowski data not shown."},{"key":"e_1_3_1_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(80)90409-2"},{"key":"e_1_3_1_9_2","doi-asserted-by":"crossref","unstructured":"Y. Ottaviano L. Gerace J. Biol. Chem. 260 624 (1985).","DOI":"10.1016\/S0021-9258(18)89778-2"},{"key":"e_1_3_1_10_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1033.2001.01901.x"},{"key":"e_1_3_1_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(90)90471-P"},{"key":"e_1_3_1_12_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.85.9.2994"},{"key":"e_1_3_1_13_2","doi-asserted-by":"publisher","DOI":"10.1007\/PL00000818"},{"key":"e_1_3_1_14_2","unstructured":"See supporting data on Science Online."},{"key":"e_1_3_1_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)68471-6"},{"key":"e_1_3_1_16_2","doi-asserted-by":"crossref","unstructured":"F. M. Jault et al. J. Virol. 69 6697 (1995).","DOI":"10.1128\/jvi.69.11.6697-6704.1995"},{"key":"e_1_3_1_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.70.12.8850-8857.1996"},{"key":"e_1_3_1_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.74.3.1355-1363.2000"},{"key":"e_1_3_1_19_2","doi-asserted-by":"crossref","unstructured":"E. S. Scott P. O'Hare J. Virol. 75 8818 (2001).","DOI":"10.1128\/JVI.75.18.8818-8830.2001"},{"key":"e_1_3_1_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.74.1.117-129.2000"},{"key":"e_1_3_1_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.4.1528"},{"key":"e_1_3_1_22_2","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.112.15.2485"},{"key":"e_1_3_1_23_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.26.15270"},{"key":"e_1_3_1_24_2","doi-asserted-by":"crossref","unstructured":"C. Laguri et al. Structure 9 503 (2001).","DOI":"10.1016\/S0969-2126(01)00611-6"},{"key":"e_1_3_1_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.70.12.8833-8849.1996"},{"key":"e_1_3_1_26_2","doi-asserted-by":"crossref","unstructured":"W. Kolanus et al. Cell 86 233 (1996).","DOI":"10.1016\/S0092-8674(00)80095-1"},{"key":"e_1_3_1_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(89)90845-3"},{"key":"e_1_3_1_28_2","first-page":"150","volume":"54","author":"Hoger T. H.","year":"1991","unstructured":"Hoger T. H., Grund C., Franke W. W., Krohne G., Eur. J. Cell Biol. 54, 150 (1991).","journal-title":"Eur. J. Cell Biol."},{"key":"e_1_3_1_29_2","unstructured":"ORFs M50 to M53 were amplified by the polymerase chain reaction (PCR) from the subcloned Hind III H fragment of the MCMV strain Smith genome [American Type Culture Collection (ATCC) VR-194] and subsequently cloned into the pCR3 (Invitrogen) plasmid by Eco RI and Xba I restriction sites. Primers for PCR were selected from the published MCMV sequence (GenBank accession number ) (24). All viral genes were expressed as immunoglobulin-tagged (cIg) (25) or FLAG-tagged fusion proteins. CV-1 cells (ATCC CCL-70) 293 cells (ATCC 1573) NIH 3T3 cells (ATCC CRL 1658) or human foreskin fibroblasts were seeded on cover slips before infection or transfection (SuperFect Qiagen) incubated 24 hours in 5% CO 2 at 37\u00b0C and then subjected to immunofluorescence."},{"key":"e_1_3_1_30_2","unstructured":"Cells were incubated with the following antibodies: anti-lamin B (Santa Cruz Biotechnology) anti-Ca-dependent PKC polyclonal rabbit serum (M. Ueffing GSF Munich Germany) anti-pp89 (26) anti-LAP2\u03b2  (R. Foisner Biocenter Vienna Austria). For the detection of M50\/p35 an affinity-purified polyclonal rabbit antiserum raised against a synthetic M50\/p35 peptide with the amino acid sequence PPGQRWGSLRKHG (G Gly; H His; K Lys; L Leu; P Pro; Q Gln; R Arg; S Ser; W Trp) was used (Eurogentec Herstal Belgium). Cells were washed and incubated with fluorescein isothiocyanate\u2013 or Texas Red\u2013conjugated secondary antibodies (Dianova Hamburg Germany). Cells subjected to DNA staining were incubated with 1 \u03bcg\/ml propidium iodide in the presence of ribonuclease A and 1 mM EDTA. Before imaging on a confocal laser scanning microscope in a sequential scan mode (Leica SP2 Bensheim Germany) cover slips were mounted with Histogel (Linaris Wertheim Germany)."},{"key":"e_1_3_1_31_2","unstructured":"Mock-infected and MCMV-infected cells were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% (v\/v) newborn calf serum (NCS) 2 mM l -glutamine penicillin (100 U\/ml) and streptomycin (100 \u03bcg\/ml). For orthophosphate labeling cells were washed in phosphate-free DMEM (Life Technologies) and incubated with [ 32 P]orthophosphate (100 \u03bcCi\/ml; Amersham Braunschweig Germany) for 3 hours (14). Cells were lysed in a buffer containing 1% Triton X-100 150 mM NaCl 20 mM tris (pH 7.5) 1 mM EDTA 0.2 mM phenylmethylsulfonyl fluoride (PMSF) 50 mM sodium fluoride 1 mM sodium vanadate and 10 \u03bcg\/ml of each of the protease inhibitors leupeptin aprotinin and soybean trypsin inhibitor. Protein concentration of lysates was determined using the BCA protein assay kit (Pierce Rockford IL) and identical amounts of protein were subjected to immunoprecipitation. An aliquot of the lysate was used for a Western blot control. For precipitation the following antibodies to lamins were used: anti-lamin A\/C (clone X67) anti-lamin B 2 (clone R29) and anti-lamin clone PKB8 (27). Samples were analyzed by 10% SDS\u2013polyacrylamide gel electrophoresis (PAGE). The PKC inhibitor Ro-31-7549 (Calbiochem San Diego CA) was used at a final concentration of 200 nM."},{"key":"e_1_3_1_32_2","unstructured":"Supported by Deutsche Forschungsgemeinschaft grants SFB 413 and SFB 455. We thank C. Atzler for technical help and M. Ueffing R. Foisner G. Gill and K. Wilson for reagents."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.1071506","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,9]],"date-time":"2024-01-09T21:58:23Z","timestamp":1704837503000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.1071506"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2002,8,2]]},"references-count":31,"journal-issue":{"issue":"5582","published-print":{"date-parts":[[2002,8,2]]}},"alternative-id":["10.1126\/science.1071506"],"URL":"https:\/\/doi.org\/10.1126\/science.1071506","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.1008505.143008","asserted-by":"object"},{"id-type":"doi","id":"10.3410\/f.1008505.107360","asserted-by":"object"}]},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2002,8,2]]}}}