DOI: 10.1128/jvi.76.10.5291-5299.2002. RNA-Binding Activity of the Rotavirus Phosphoprotein NSP5 Includes Affinity for Double-Stranded RNA

RNA-Binding Activity of the Rotavirus Phosphoprotein NSP5 Includes Affinity for Double-Stranded RNA

10.1128/jvi.76.10.5291-5299.2002

Crossref journal-article

American Society for Microbiology

Journal of Virology (235)

Abstract

ABSTRACT Phosphoprotein NSP5 is a component of replication intermediates that catalyze the synthesis of the segmented double-stranded RNA (dsRNA) rotavirus genome. To study the role of the protein in viral replication, His-tagged NSP5 was expressed in bacteria and purified by affinity chromatography. In vitro phosphorylation assays showed that NSP5 alone contains minimal autokinase activity but undergoes hyperphosphorylation when combined with the NTPase and helix-destabilizing protein NSP2. Hence, NSP2 mediates the hyperphosphorylation of NSP5 in the absence of other viral or cellular proteins. RNA-binding assays demonstrated that NSP5 has unique nonspecific RNA-binding activity, recognizing single-stranded RNA and dsRNA with similar affinities. The possible functions of the RNA-binding activities of NSP5 are to cooperate with NSP2 in the destabilization of RNA secondary structures and in the packaging of RNA and/or to prevent the interferon-induced dsRNA-dependent activation of the protein kinase PKR.

Bibliography

Vende, P., Taraporewala, Z. F., & Patton, J. T. (2002). RNA-Binding Activity of the Rotavirus Phosphoprotein NSP5 Includes Affinity for Double-Stranded RNA. Journal of Virology, 76(10), 5291â5299.

Authors 3

Patrice Vende (first)
Zenobia F. Taraporewala (additional)
John T. Patton (additional)

References 51 Referenced 50

10.1099/0022-1317-79-11-2679
10.1099/0022-1317-77-9-2059
10.1128/jvi.70.2.985-991.1996
10.1006/jsbi.2001.4352
10.1128/jvi.71.1.138-144.1997
10.1128/JVI.72.8.6398-6405.1998
10.1128/jvi.58.2.561-568.1986
10.1006/viro.1999.0029
10.1128/jvi.68.11.7030-7039.1994
10.1016/S1357-2725(96)00169-0
Estes, M. K. 2001. Rotaviruses and their replication, p. 1747-1785. In D. M. Knipe, P. M. Howley, et al. (ed.), Fields virology, 4th ed. Lippincott Williams & Wilkins, Philadelphia, Pa. / Fields virology (2001)
10.1099/0022-1317-80-2-333
10.1016/S0968-0004(00)01580-2
10.1016/0042-6822(89)90204-3
10.1016/0042-6822(91)90642-O
10.1007/s007050050347
10.1128/jvi.68.6.3990-4000.1994
Kapikian, A. Z., Y. Hoshino, and R. M. Chanock. 2001. Rotaviruses, p. 1787-1833. In D. M. Knipe, P. M. Howley, et al. (ed.), Fields virology, 4th ed. Lippincott Williams & Wilkins, Philadelphia, Pa. / Fields virology (2001)
10.1006/viro.1994.1402
10.1016/0042-6822(92)90245-K
10.1099/0022-1317-75-12-3423
10.1128/jvi.68.6.3821-3829.1994
Lawton, J. A., M. K. Estes, and B. V. V. Prasad. 1997. Three-dimensional visualization of mRNA release from actively transcribing rotavirus particles. Nat. Struct. Biol. 2 : 118-121. / Nat. Struct. Biol. (1997)
10.1128/jvi.71.10.7353-7360.1997
10.1016/0042-6822(92)90736-9
10.1016/0042-6822(91)90495-W
10.1016/S0065-3527(08)60355-3
10.1093/nar/16.13.6244
10.1093/emboj/20.5.979
10.1128/jvi.70.11.7940-7947.1996
10.1128/JVI.73.2.1382-1391.1999
10.1099/0022-1317-71-5-1087
10.1016/S1067-2389(06)80056-7
10.1006/viro.2000.0645
10.1128/jvi.70.6.3961-3971.1996
Patton, J. T., Z. Taraporewala, V. Chizhikov, and D. Chen. 1999. Virus replication. Methods Mol. Med. 34 : 33-66. / Methods Mol. Med. (1999)
10.1016/0168-1702(84)90069-8
10.1002/j.1460-2075.1994.tb06734.x
10.1128/jvi.71.1.34-41.1997
10.1016/0022-2836(88)90313-0
10.1128/jvi.69.9.5502-5505.1995
10.1021/bi9824036
10.1128/mcb.8.1.273-283.1988
Schuck, P., Z. Taraporewala, P. McPhie, and J. T. Patton. 2000. Rotavirus nonstructural protein NSP2 self-assembles into octamers that undergo ligand-induced conformational changes. J. Biol. Chem. 276 : 9679-9687. / J. Biol. Chem. (2000)
10.1128/jvi.71.10.7220-7226.1997
10.1128/JVI.75.10.4519-4527.2001
10.1128/JVI.73.12.9934-9943.1999
10.1006/viro.2000.0764
10.1099/0022-1317-81-3-821
10.1128/jvi.65.7.3964-3967.1991
10.1006/viro.1997.8664

Dates

Type	When
Created	23 years ago (July 27, 2002, 6:09 a.m.)
Deposited	3 years, 5 months ago (March 5, 2022, 1:53 a.m.)
Indexed	1 month, 3 weeks ago (June 29, 2025, 11:01 a.m.)
Issued	23 years, 3 months ago (May 15, 2002)
Published	23 years, 3 months ago (May 15, 2002)
Published Print	23 years, 3 months ago (May 15, 2002)

Funders 0

None

BibTeX

@article{Vende_2002, title={RNA-Binding Activity of the Rotavirus Phosphoprotein NSP5 Includes Affinity for Double-Stranded RNA}, volume={76}, ISSN={1098-5514}, url={http://dx.doi.org/10.1128/jvi.76.10.5291-5299.2002}, DOI={10.1128/jvi.76.10.5291-5299.2002}, number={10}, journal={Journal of Virology}, publisher={American Society for Microbiology}, author={Vende, Patrice and Taraporewala, Zenobia F. and Patton, John T.}, year={2002}, month=may, pages={5291–5299} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        6,
        29
      ]
    ],
    "date-time": "2025-06-29T15:01:41Z",
    "timestamp": 1751209301419
  },
  "reference-count": 51,
  "publisher": "American Society for Microbiology",
  "issue": "10",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2002,
            5,
            15
          ]
        ],
        "date-time": "2002-05-15T00:00:00Z",
        "timestamp": 1021420800000
      },
      "content-version": "tdm",
      "delay-in-days": 0,
      "URL": "https://journals.asm.org/non-commercial-tdm-license"
    }
  ],
  "content-domain": {
    "domain": [
      "journals.asm.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        2002,
        5,
        15
      ]
    ]
  },
  "abstract": "<jats:title>ABSTRACT</jats:title>\n          <jats:p>Phosphoprotein NSP5 is a component of replication intermediates that catalyze the synthesis of the segmented double-stranded RNA (dsRNA) rotavirus genome. To study the role of the protein in viral replication, His-tagged NSP5 was expressed in bacteria and purified by affinity chromatography. In vitro phosphorylation assays showed that NSP5 alone contains minimal autokinase activity but undergoes hyperphosphorylation when combined with the NTPase and helix-destabilizing protein NSP2. Hence, NSP2 mediates the hyperphosphorylation of NSP5 in the absence of other viral or cellular proteins. RNA-binding assays demonstrated that NSP5 has unique nonspecific RNA-binding activity, recognizing single-stranded RNA and dsRNA with similar affinities. The possible functions of the RNA-binding activities of NSP5 are to cooperate with NSP2 in the destabilization of RNA secondary structures and in the packaging of RNA and/or to prevent the interferon-induced dsRNA-dependent activation of the protein kinase PKR.</jats:p>",
  "DOI": "10.1128/jvi.76.10.5291-5299.2002",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        27
      ]
    ],
    "date-time": "2002-07-27T10:09:29Z",
    "timestamp": 1027764569000
  },
  "page": "5291-5299",
  "update-policy": "http://dx.doi.org/10.1128/asmj-crossmark-policy-page",
  "source": "Crossref",
  "is-referenced-by-count": 50,
  "title": "RNA-Binding Activity of the Rotavirus Phosphoprotein NSP5 Includes Affinity for Double-Stranded RNA",
  "prefix": "10.1128",
  "volume": "76",
  "author": [
    {
      "given": "Patrice",
      "family": "Vende",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892"
        },
        {
          "name": "Laboratoire de Virologie et Immunologie Mole\u0301culaires INRA, CRJJ, Domaine de Vilvert, 78352 Jouy-en-Josas Cedex, France"
        }
      ]
    },
    {
      "given": "Zenobia F.",
      "family": "Taraporewala",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892"
        }
      ]
    },
    {
      "given": "John T.",
      "family": "Patton",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892"
        }
      ]
    }
  ],
  "member": "235",
  "reference": [
    {
      "key": "e_1_3_2_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/0022-1317-79-11-2679"
    },
    {
      "key": "e_1_3_2_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/0022-1317-77-9-2059"
    },
    {
      "key": "e_1_3_2_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.70.2.985-991.1996"
    },
    {
      "key": "e_1_3_2_5_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jsbi.2001.4352"
    },
    {
      "key": "e_1_3_2_6_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.71.1.138-144.1997"
    },
    {
      "key": "e_1_3_2_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JVI.72.8.6398-6405.1998"
    },
    {
      "key": "e_1_3_2_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.58.2.561-568.1986"
    },
    {
      "key": "e_1_3_2_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/viro.1999.0029"
    },
    {
      "key": "e_1_3_2_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.68.11.7030-7039.1994"
    },
    {
      "key": "e_1_3_2_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1357-2725(96)00169-0"
    },
    {
      "key": "e_1_3_2_12_2",
      "first-page": "1747",
      "year": "2001",
      "unstructured": "Estes, M. K. 2001. Rotaviruses and their replication, p. 1747-1785. In D. M. Knipe, P. M. Howley, et al. (ed.), Fields virology, 4th ed. Lippincott Williams & Wilkins, Philadelphia, Pa.",
      "journal-title": "Fields virology"
    },
    {
      "key": "e_1_3_2_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/0022-1317-80-2-333"
    },
    {
      "key": "e_1_3_2_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0968-0004(00)01580-2"
    },
    {
      "key": "e_1_3_2_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0042-6822(89)90204-3"
    },
    {
      "key": "e_1_3_2_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0042-6822(91)90642-O"
    },
    {
      "key": "e_1_3_2_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/s007050050347"
    },
    {
      "key": "e_1_3_2_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.68.6.3990-4000.1994"
    },
    {
      "key": "e_1_3_2_19_2",
      "first-page": "1787",
      "year": "2001",
      "unstructured": "Kapikian, A. Z., Y. Hoshino, and R. M. Chanock. 2001. Rotaviruses, p. 1787-1833. In D. M. Knipe, P. M. Howley, et al. (ed.), Fields virology, 4th ed. Lippincott Williams & Wilkins, Philadelphia, Pa.",
      "journal-title": "Fields virology"
    },
    {
      "key": "e_1_3_2_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/viro.1994.1402"
    },
    {
      "key": "e_1_3_2_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0042-6822(92)90245-K"
    },
    {
      "key": "e_1_3_2_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/0022-1317-75-12-3423"
    },
    {
      "key": "e_1_3_2_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.68.6.3821-3829.1994"
    },
    {
      "key": "e_1_3_2_24_2",
      "first-page": "118",
      "volume": "2",
      "year": "1997",
      "unstructured": "Lawton, J. A., M. K. Estes, and B. V. V. Prasad. 1997. Three-dimensional visualization of mRNA release from actively transcribing rotavirus particles. Nat. Struct. Biol. 2 : 118-121.",
      "journal-title": "Nat. Struct. Biol."
    },
    {
      "key": "e_1_3_2_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.71.10.7353-7360.1997"
    },
    {
      "key": "e_1_3_2_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0042-6822(92)90736-9"
    },
    {
      "key": "e_1_3_2_27_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0042-6822(91)90495-W"
    },
    {
      "key": "e_1_3_2_28_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0065-3527(08)60355-3"
    },
    {
      "key": "e_1_3_2_29_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/16.13.6244"
    },
    {
      "key": "e_1_3_2_30_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/20.5.979"
    },
    {
      "key": "e_1_3_2_31_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.70.11.7940-7947.1996"
    },
    {
      "key": "e_1_3_2_32_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JVI.73.2.1382-1391.1999"
    },
    {
      "key": "e_1_3_2_33_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/0022-1317-71-5-1087"
    },
    {
      "key": "e_1_3_2_34_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1067-2389(06)80056-7"
    },
    {
      "key": "e_1_3_2_35_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/viro.2000.0645"
    },
    {
      "key": "e_1_3_2_36_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.70.6.3961-3971.1996"
    },
    {
      "key": "e_1_3_2_37_2",
      "first-page": "33",
      "volume": "34",
      "year": "1999",
      "unstructured": "Patton, J. T., Z. Taraporewala, V. Chizhikov, and D. Chen. 1999. Virus replication. Methods Mol. Med. 34 : 33-66.",
      "journal-title": "Methods Mol. Med."
    },
    {
      "key": "e_1_3_2_38_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0168-1702(84)90069-8"
    },
    {
      "key": "e_1_3_2_39_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1994.tb06734.x"
    },
    {
      "key": "e_1_3_2_40_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.71.1.34-41.1997"
    },
    {
      "key": "e_1_3_2_41_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0022-2836(88)90313-0"
    },
    {
      "key": "e_1_3_2_42_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.69.9.5502-5505.1995"
    },
    {
      "key": "e_1_3_2_43_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi9824036"
    },
    {
      "key": "e_1_3_2_44_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/mcb.8.1.273-283.1988"
    },
    {
      "key": "e_1_3_2_45_2",
      "first-page": "9679",
      "volume": "276",
      "year": "2000",
      "unstructured": "Schuck, P., Z. Taraporewala, P. McPhie, and J. T. Patton. 2000. Rotavirus nonstructural protein NSP2 self-assembles into octamers that undergo ligand-induced conformational changes. J. Biol. Chem. 276 : 9679-9687.",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_3_2_46_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.71.10.7220-7226.1997"
    },
    {
      "key": "e_1_3_2_47_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JVI.75.10.4519-4527.2001"
    },
    {
      "key": "e_1_3_2_48_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JVI.73.12.9934-9943.1999"
    },
    {
      "key": "e_1_3_2_49_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/viro.2000.0764"
    },
    {
      "key": "e_1_3_2_50_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/0022-1317-81-3-821"
    },
    {
      "key": "e_1_3_2_51_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.65.7.3964-3967.1991"
    },
    {
      "key": "e_1_3_2_52_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/viro.1997.8664"
    }
  ],
  "container-title": "Journal of Virology",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://journals.asm.org/doi/pdf/10.1128/JVI.76.10.5291-5299.2002",
      "content-type": "application/pdf",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://journals.asm.org/doi/pdf/10.1128/JVI.76.10.5291-5299.2002",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2022,
        3,
        5
      ]
    ],
    "date-time": "2022-03-05T06:53:37Z",
    "timestamp": 1646463217000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://journals.asm.org/doi/10.1128/JVI.76.10.5291-5299.2002"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2002,
        5,
        15
      ]
    ]
  },
  "references-count": 51,
  "journal-issue": {
    "issue": "10",
    "published-print": {
      "date-parts": [
        [
          2002,
          5,
          15
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1128/JVI.76.10.5291-5299.2002"
  ],
  "URL": "http://dx.doi.org/10.1128/JVI.76.10.5291-5299.2002",
  "relation": {},
  "ISSN": [
    "0022-538X",
    "1098-5514"
  ],
  "subject": [],
  "container-title-short": "J Virol",
  "published": {
    "date-parts": [
      [
        2002,
        5,
        15
      ]
    ]
  },
  "assertion": [
    {
      "value": "2001-11-14",
      "order": 0,
      "name": "received",
      "label": "Received",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "2002-02-20",
      "order": 1,
      "name": "accepted",
      "label": "Accepted",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "2002-05-15",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}