Intracellular localisation of dengue-2 RNA in mosquito cell culture using electron microscopic in situ hybridisation

被引:41
作者
Grief, C
Galler, R
Côrtes, LMC
Barth, OM
机构
[1] FIOCRUZ, Inst Oswaldo Cruz, Dept Virol, BR-21045900 Rio De Janeiro, Brazil
[2] FIOCRUZ, Inst Oswaldo Cruz, Dept Biochem & Mol Biol, BR-21045900 Rio De Janeiro, Brazil
关键词
D O I
10.1007/s007050050247
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Non-isotopic in situ hybridisation was used at the electron microscope level to determine the localisation of viral RNA in dengue-2 infected mosquito cells at 14, 24, 48 and 72h post-infection. In situ hybridisation was carried out on sections of dengue-2 infected mosquito cells using a digoxigenin-labelled DNA probe to the envelope protein gene sequence of the virus. Viral RNA was consistently localised over the rough endoplasmic reticulum and the virus-induced smooth membrane structures which form within the endoplasmic reticulum. During the later stages of infection electron-dense areas were observed to develop in close proximity to the smooth membrane structures. Electron microscopic in situ hybridisation showed that these denser areas contained both viral RNA and virus particles. Our results show that in dengue-2 infected mosquito cells the smooth membrane structures are an important site for the concentration of dengue viral RNA and its possible subsequent encapsidation into virus particles.
引用
收藏
页码:2347 / 2357
页数:11
相关论文
共 20 条
[1]   GENERATION OF DIGOXIGENIN-LABELED DOUBLE-STRANDED AND SINGLE-STRANDED PROBES USING THE POLYMERASE CHAIN-REACTION [J].
AN, SF ;
FRANKLIN, D ;
FLEMING, KA .
MOLECULAR AND CELLULAR PROBES, 1992, 6 (03) :193-200
[2]   REPLICATION OF DENGUE VIRUSES IN MOSQUITO CELL-CULTURES - A MODEL FROM ULTRASTRUCTURAL OBSERVATIONS [J].
BARTH, OM .
MEMORIAS DO INSTITUTO OSWALDO CRUZ, 1992, 87 (04) :565-574
[3]   STRUCTURAL AND FUNCTIONAL-CHARACTERIZATION OF THE POLIOVIRUS REPLICATION COMPLEX [J].
BIENZ, K ;
EGGER, D ;
PFISTER, T ;
TROXLER, M .
JOURNAL OF VIROLOGY, 1992, 66 (05) :2740-2747
[4]   ASSOCIATION OF POLIOVIRAL PROTEINS OF THE P2-GENOMIC REGION WITH THE VIRAL REPLICATION COMPLEX AND VIRUS-INDUCED MEMBRANE SYNTHESIS AS VISUALIZED BY ELECTRON-MICROSCOPIC IMMUNOCYTOCHEMISTRY AND AUTORADIOGRAPHY [J].
BIENZ, K ;
EGGER, D ;
PASAMONTES, L .
VIROLOGY, 1987, 160 (01) :220-226
[5]   CHARACTERIZATION OF POLIOVIRUS-SPECIFIC STRUCTURES ASSOCIATED WITH CYTOPLASMIC MEMBRANES [J].
CALIGUIRI, LA ;
TAMM, I .
VIROLOGY, 1970, 42 (01) :112-+
[6]   MOLECULAR AND ULTRASTRUCTURAL ANALYSIS OF HEAVY MEMBRANE-FRACTIONS ASSOCIATED WITH THE REPLICATION OF KUNJIN VIRUS-RNA [J].
CHU, PWG ;
WESTAWAY, EG .
ARCHIVES OF VIROLOGY, 1992, 125 (1-4) :177-191
[7]   COMPARISON OF REPLICATION COMPLEXES SYNTHESIZING POLIOVIRUS RNA [J].
ETCHISON, D ;
EHRENFELD, E .
VIROLOGY, 1981, 111 (01) :33-46
[8]  
FIELDS BN, 1990, CIROLOGY
[9]   THE ASSEMBLY OF HIV WITHIN THE GOLGI-APPARATUS AND GOLGI-DERIVED VESICLES OF JM CELL SYNCYTIA [J].
GRIEF, C ;
FARRAR, GH ;
KENT, KA ;
BERGER, EG .
AIDS, 1991, 5 (12) :1433-1439
[10]   NUCLEOTIDE-SEQUENCE OF DENGUE-2 RNA AND COMPARISON OF THE ENCODED PROTEINS WITH THOSE OF OTHER FLAVIVIRUSES [J].
HAHN, YS ;
GALLER, R ;
HUNKAPILLER, T ;
DALRYMPLE, JM ;
STRAUSS, JH ;
STRAUSS, EG .
VIROLOGY, 1988, 162 (01) :167-180