Abstract
The transcription and replication of influenza RNA can be studied in vitro by the reconstitution of functional ribonucleoprotein (RNP) complex from viral core proteins including the RNA polymerase (complex of three P protein subunits) and nucleoprotein (NP), and model templates. Here, two different core protein preparations, one based on CsCl centrifugation (CS enzyme) and the other on micrococcal nuclease treatment of viral cores (MN enzyme), were compared side-by-side. Short model RNA templates and their 3′-half molecules of both viral RNA (vRNA) and complementary RNA (cRNA) senses were reconstituted with the core protein preparations in parallel, and RNA polymerase activity was tested either in the presence or absence of ApG or globin mRNA as primers. Both enzyme preparations were active in the syntheses of short vRNA and cRNA transcripts using ApG as a primer, although the synthesis of cRNA was 2-10-fold higher (depending on the template used) than the synthesis of vRNA. The MN enzyme, however, was more active per weight of total protein than the CS enzyme, probably because of its higher content of RNA polymerase. Both enzymes failed to show primer-independent synthesis of vRNA. The differences observed in the synthesis of short transcripts using globin mRNA as a primer are discussed.
| Original language | English |
|---|---|
| Pages (from-to) | 496-499 |
| Number of pages | 4 |
| Journal | Journal of Biochemistry |
| Volume | 111 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 1992 Jan 1 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
Cite this
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Comparison of two reconstituted systems for In Vitro transcription and replication of influenza virus. / Seong, Baik L.; Kobayashi, Makoto; Nagata, Kyosuke; Brownlee, George G.; Ishihama, Akira.
In: Journal of Biochemistry, Vol. 111, No. 4, 01.01.1992, p. 496-499.Research output: Contribution to journal › Article
TY - JOUR
T1 - Comparison of two reconstituted systems for In Vitro transcription and replication of influenza virus
AU - Seong, Baik L.
AU - Kobayashi, Makoto
AU - Nagata, Kyosuke
AU - Brownlee, George G.
AU - Ishihama, Akira
PY - 1992/1/1
Y1 - 1992/1/1
N2 - The transcription and replication of influenza RNA can be studied in vitro by the reconstitution of functional ribonucleoprotein (RNP) complex from viral core proteins including the RNA polymerase (complex of three P protein subunits) and nucleoprotein (NP), and model templates. Here, two different core protein preparations, one based on CsCl centrifugation (CS enzyme) and the other on micrococcal nuclease treatment of viral cores (MN enzyme), were compared side-by-side. Short model RNA templates and their 3′-half molecules of both viral RNA (vRNA) and complementary RNA (cRNA) senses were reconstituted with the core protein preparations in parallel, and RNA polymerase activity was tested either in the presence or absence of ApG or globin mRNA as primers. Both enzyme preparations were active in the syntheses of short vRNA and cRNA transcripts using ApG as a primer, although the synthesis of cRNA was 2-10-fold higher (depending on the template used) than the synthesis of vRNA. The MN enzyme, however, was more active per weight of total protein than the CS enzyme, probably because of its higher content of RNA polymerase. Both enzymes failed to show primer-independent synthesis of vRNA. The differences observed in the synthesis of short transcripts using globin mRNA as a primer are discussed.
AB - The transcription and replication of influenza RNA can be studied in vitro by the reconstitution of functional ribonucleoprotein (RNP) complex from viral core proteins including the RNA polymerase (complex of three P protein subunits) and nucleoprotein (NP), and model templates. Here, two different core protein preparations, one based on CsCl centrifugation (CS enzyme) and the other on micrococcal nuclease treatment of viral cores (MN enzyme), were compared side-by-side. Short model RNA templates and their 3′-half molecules of both viral RNA (vRNA) and complementary RNA (cRNA) senses were reconstituted with the core protein preparations in parallel, and RNA polymerase activity was tested either in the presence or absence of ApG or globin mRNA as primers. Both enzyme preparations were active in the syntheses of short vRNA and cRNA transcripts using ApG as a primer, although the synthesis of cRNA was 2-10-fold higher (depending on the template used) than the synthesis of vRNA. The MN enzyme, however, was more active per weight of total protein than the CS enzyme, probably because of its higher content of RNA polymerase. Both enzymes failed to show primer-independent synthesis of vRNA. The differences observed in the synthesis of short transcripts using globin mRNA as a primer are discussed.
UR - http://www.scopus.com/inward/record.url?scp=0026596569&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026596569&partnerID=8YFLogxK
U2 - 10.1093/oxfordjournals.jbchem.a123786
DO - 10.1093/oxfordjournals.jbchem.a123786
M3 - Article
C2 - 1618740
AN - SCOPUS:0026596569
VL - 111
SP - 496
EP - 499
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 4
ER -