The RNA binding domain of the hantaan virus N protein maps to a central, conserved region
The nucleocapsid (N) protein of hantaviruses encapsidates both viral genomic and antigenomic RNAs, although only the genomic viral RNA (vRNA) is packaged into virions. To define the domain within the Hantaan virus (HTNV) N protein that mediates these interactions, 14 N- and C-terminal deletion constructs were cloned into a bacterial expression vector, expressed, and purified to homogeneity. Each protein was examined for its ability to bind the HTNV S segment vRNA with filter binding and gel electrophoretic mobility shift assays. These studies mapped a minimal region within the HTNV N protein (amino acids 175 to 217) that bound vRNA. Sequence alignments made from several hantavirus N protein sequences showed that the region identified has a 58% identity and an 86% similarity among these amino acid sequences. Two peptides corresponding to amino acids 175 to 196 (N1) and 197 to 218 (N2) were synthesized. The RNA binding of each peptide was measured by filter binding and competition analysis. Three oligoribonucleotides were used to measure binding affinity and assess specificity. The N2 peptide contained the major RNA binding determinants, while the N1 peptide, when mixed with N2, contributed to the specificity of vRNA recognition.
Medienart: |
Artikel |
---|
Erscheinungsjahr: |
2002 |
---|---|
Erschienen: |
2002 |
Enthalten in: |
Zur Gesamtaufnahme - volume:76 |
---|---|
Enthalten in: |
Journal of virology - 76(2002), 7 vom: 15. Apr., Seite 3301-8 |
Sprache: |
Englisch |
---|
Beteiligte Personen: |
Xu, Xiaolin [VerfasserIn] |
---|
Themen: |
Capsid Proteins |
---|
Anmerkungen: |
Date Completed 19.04.2002 Date Revised 08.05.2019 published: Print Citation Status MEDLINE |
---|
Förderinstitution / Projekttitel: |
|
---|
PPN (Katalog-ID): |
NLM117688339 |
---|
LEADER | 01000naa a22002652 4500 | ||
---|---|---|---|
001 | NLM117688339 | ||
003 | DE-627 | ||
005 | 20231222181100.0 | ||
007 | tu | ||
008 | 231222s2002 xx ||||| 00| ||eng c | ||
028 | 5 | 2 | |a pubmed24n0393.xml |
035 | |a (DE-627)NLM117688339 | ||
035 | |a (NLM)11884555 | ||
040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
041 | |a eng | ||
100 | 1 | |a Xu, Xiaolin |e verfasserin |4 aut | |
245 | 1 | 4 | |a The RNA binding domain of the hantaan virus N protein maps to a central, conserved region |
264 | 1 | |c 2002 | |
336 | |a Text |b txt |2 rdacontent | ||
337 | |a ohne Hilfsmittel zu benutzen |b n |2 rdamedia | ||
338 | |a Band |b nc |2 rdacarrier | ||
500 | |a Date Completed 19.04.2002 | ||
500 | |a Date Revised 08.05.2019 | ||
500 | |a published: Print | ||
500 | |a Citation Status MEDLINE | ||
520 | |a The nucleocapsid (N) protein of hantaviruses encapsidates both viral genomic and antigenomic RNAs, although only the genomic viral RNA (vRNA) is packaged into virions. To define the domain within the Hantaan virus (HTNV) N protein that mediates these interactions, 14 N- and C-terminal deletion constructs were cloned into a bacterial expression vector, expressed, and purified to homogeneity. Each protein was examined for its ability to bind the HTNV S segment vRNA with filter binding and gel electrophoretic mobility shift assays. These studies mapped a minimal region within the HTNV N protein (amino acids 175 to 217) that bound vRNA. Sequence alignments made from several hantavirus N protein sequences showed that the region identified has a 58% identity and an 86% similarity among these amino acid sequences. Two peptides corresponding to amino acids 175 to 196 (N1) and 197 to 218 (N2) were synthesized. The RNA binding of each peptide was measured by filter binding and competition analysis. Three oligoribonucleotides were used to measure binding affinity and assess specificity. The N2 peptide contained the major RNA binding determinants, while the N1 peptide, when mixed with N2, contributed to the specificity of vRNA recognition | ||
650 | 4 | |a Comparative Study | |
650 | 4 | |a Journal Article | |
650 | 4 | |a Research Support, U.S. Gov't, P.H.S. | |
650 | 7 | |a Capsid Proteins |2 NLM | |
650 | 7 | |a Peptides |2 NLM | |
650 | 7 | |a RNA, Viral |2 NLM | |
650 | 7 | |a RNA-Binding Proteins |2 NLM | |
650 | 7 | |a Viral Core Proteins |2 NLM | |
650 | 7 | |a nucleocapsid protein, Hantaan virus |2 NLM | |
700 | 1 | |a Severson, William |e verfasserin |4 aut | |
700 | 1 | |a Villegas, Noah |e verfasserin |4 aut | |
700 | 1 | |a Schmaljohn, Connie S |e verfasserin |4 aut | |
700 | 1 | |a Jonsson, Colleen B |e verfasserin |4 aut | |
773 | 0 | 8 | |i Enthalten in |t Journal of virology |d 1967 |g 76(2002), 7 vom: 15. Apr., Seite 3301-8 |w (DE-627)NLM000006866 |x 1098-5514 |7 nnns |
773 | 1 | 8 | |g volume:76 |g year:2002 |g number:7 |g day:15 |g month:04 |g pages:3301-8 |
912 | |a GBV_USEFLAG_A | ||
912 | |a GBV_NLM | ||
951 | |a AR | ||
952 | |d 76 |j 2002 |e 7 |b 15 |c 04 |h 3301-8 |