eprintid: 8381
rev_number: 14
eprint_status: archive
userid: 1
dir: disk0/00/00/83/81
datestamp: 2008-04-30 17:27:21
lastmod: 2016-01-01 12:10:32
status_changed: 2012-08-14 15:25:16
type: article
metadata_visibility: show
creators_name: Sehgal, Pravinkumar B.
creators_name: Dobberstein, Bernhard
creators_name: Tamm, Igor
title: Interferon messenger RNA content of human fibroblasts during induction, shutoff, and superinduction of interferon production
ispublished: pub
subjects: 570
divisions: 706000
keywords: human diploid fibroblast cells , poly(I)poly(C) , McAuslan-Tomkins translational repressor hypothesis , 5,6-dichloro-l-l-Dribofuranosylbenzimidazole
abstract: Translation of injected mRNA in oocytes of Xenopus laevis has been used as a highly sensitive and quantitative assay for interferon mRNA. Injection into oocytes of polyadenylylated RNA extracted from poly(I)poly(C)induced human diploid fibroblasts (FS-4) leads to the synthesis of biologically active human fibroblast interferon over a period of 24-32 hr. There is a linear relationship between the amount of mRNA injected and the interferon yield obtained over a range of 1-20 ng of injected RNA. Injection of 40-80 ng of mRNA into each of 15 oocytes, homogenized in 0.3 ml of incubation medium, gave a titer of 128-256 interferon reference units/ml of homogenate. FS-4 cells at the peak of interferon production-i.e., approximately 2.5 hr after the beginning of induction with poly(I)poly(C-gave mRNA that yielded 24-48 interferon reference units/ml in the oocyte assay (30 ng of RNA injected per oocyte). An equivalent amount of mRNA from FS-4 cells in the shutoff phase, approximately 6 hr after induction, gave <4 interferon reference units/ml. In contrast, mRNA extracted from FS-4 cells that had been induced and maintained in the presence of 40 uM 5,6-dichloro-1-j-o-ribofuranosylbenzimidazole for 6 hr produced 64-128 interferon reference units/ml. Polyadenylylated RNA obtained from uninduced FS-4 cells did not lead to detectable interferon synthesis (<4 interferon reference units/ml). These data provide a direct verification of the hypothesis that the shutoff of interferon production in FS-4 cells involves a regulatory event leading to the posttranscriptional inactivation or degradation of interferon mRNA. Because the inactivating mechanism is sensitive to inhibition by 5,6-dichloro- 1--D-ribofuranosylbenzimidazole, a selective inhibitor of nuclear heterogeneous RNA and mRNA synthesis, it is likely that synthesis of an RNA molecule is necessary for the shutoff of interferon production.
abstract_translated_lang: eng
date: 1977
date_type: published
id_scheme: DOI
id_number: 10.11588/heidok.00008381
schriftenreihe_cluster_id: sr-2
schriftenreihe_order: 006
ppn_swb: 1349723258
own_urn: urn:nbn:de:bsz:16-opus-83817
language: eng
bibsort: SEHGALPRAVINTERFERON1977
full_text_status: public
publication: Proc. Natl. Acad. Sci. USA
volume: 74
number: 8
pagerange: 3409-3413
citation:   Sehgal, Pravinkumar B. ; Dobberstein, Bernhard ; Tamm, Igor  (1977) Interferon messenger RNA content of human fibroblasts during induction, shutoff, and superinduction of interferon production.  Proc. Natl. Acad. Sci. USA, 74 (8).  pp. 3409-3413.      
document_url: https://archiv.ub.uni-heidelberg.de/volltextserver/8381/1/Sehgal_1977_PNAS.pdf