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Nucleocapsid formation and RNA synthesis of Marburg virus is dependent on two coiled coil motifs in the nucleoprotein.

DiCarlo A, Möller P, Lander A, Kolesnikova L, Becker S - Virol. J. (2007)

Bottom Line: In the present study, a conserved coiled coil motif in the central part of MARV NP was shown to be an important element for the interactions of NP with itself and VP35, the viral polymerase cofactor.Additionally, the coiled coil motif was essential for the formation of NP-induced intracellular inclusions and for the function of NP in the process of transcription and replication of viral RNA in a minigenome system.The coiled coil motif is bipartite, constituted by two coiled coils which are separated by a flexible linker.

View Article: PubMed Central - HTML - PubMed

Affiliation: Philipps-Universität Marburg, Institut für Virologie, Hans Meerwein-Str, 2, 35032 Marburg, Germany. andrea.dicarlo@promega.com

ABSTRACT
The nucleoprotein (NP) of Marburg virus (MARV) is responsible for the encapsidation of viral genomic RNA and the formation of the helical nucleocapsid precursors that accumulate in intracellular inclusions in infected cells. To form the large helical MARV nucleocapsid, NP needs to interact with itself and the viral proteins VP30, VP35 and L, which are also part of the MARV nucleocapsid. In the present study, a conserved coiled coil motif in the central part of MARV NP was shown to be an important element for the interactions of NP with itself and VP35, the viral polymerase cofactor. Additionally, the coiled coil motif was essential for the formation of NP-induced intracellular inclusions and for the function of NP in the process of transcription and replication of viral RNA in a minigenome system. Transfer of the coiled coil motif to a reporter protein was sufficient to mediate interaction of the constructed fusion protein with the N-terminus of NP. The coiled coil motif is bipartite, constituted by two coiled coils which are separated by a flexible linker.

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Coiled coil motif in Zaire Ebola virus NP (A) In silico analysis of EBOV NP predicted one coiled coil motifs at aa position 333 to 367. (B) Alignment of the coiled coil regions in NP of filoviruses. C1: coiled coil motif 1, C2: coiled coil motif 2. Linker: Sequence without coiled coil prediction. a, d: Key positions in the heptad repeats of MARV NP. Large boxes: conserved amino acids at the coiled coil key positions. Small boxes: Key positions in MARV NP without conservation in Ebola virus NP.
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Figure 5: Coiled coil motif in Zaire Ebola virus NP (A) In silico analysis of EBOV NP predicted one coiled coil motifs at aa position 333 to 367. (B) Alignment of the coiled coil regions in NP of filoviruses. C1: coiled coil motif 1, C2: coiled coil motif 2. Linker: Sequence without coiled coil prediction. a, d: Key positions in the heptad repeats of MARV NP. Large boxes: conserved amino acids at the coiled coil key positions. Small boxes: Key positions in MARV NP without conservation in Ebola virus NP.

Mentions: We have recently reported that homooligomerization of VP35, the polymerase cofactor, is mediated by a coiled coil motif which leads to the formation of tetramers [10]. Here we show that the predicted coiled coil domains in NP, the major MARV nucleocapsid protein, play an essential role in the formation of NP-NP oligomers and the formation of inclusion bodies which contain preformed NP-induced nucleocapsid-like structures [8]. The nucleoprotein of Ebola virus also contains a region, encompassing amino acids 334 to 363, which has a very high probability to form a coiled coil structure (Fig. 5A). The predicted coiled coil in Ebola virus NP corresponds to the predicted coiled coil C1 of MARV NP. Seven of the nine key residues of C1 are conserved between MARV and ZEBOV (Fig. 5B). Interestingly, the second coiled coil predicted in MARV NP (amino acid positions 371 – 400) is less conserved in Ebola virus NP (Fig. 5B).


Nucleocapsid formation and RNA synthesis of Marburg virus is dependent on two coiled coil motifs in the nucleoprotein.

DiCarlo A, Möller P, Lander A, Kolesnikova L, Becker S - Virol. J. (2007)

Coiled coil motif in Zaire Ebola virus NP (A) In silico analysis of EBOV NP predicted one coiled coil motifs at aa position 333 to 367. (B) Alignment of the coiled coil regions in NP of filoviruses. C1: coiled coil motif 1, C2: coiled coil motif 2. Linker: Sequence without coiled coil prediction. a, d: Key positions in the heptad repeats of MARV NP. Large boxes: conserved amino acids at the coiled coil key positions. Small boxes: Key positions in MARV NP without conservation in Ebola virus NP.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2170442&req=5

Figure 5: Coiled coil motif in Zaire Ebola virus NP (A) In silico analysis of EBOV NP predicted one coiled coil motifs at aa position 333 to 367. (B) Alignment of the coiled coil regions in NP of filoviruses. C1: coiled coil motif 1, C2: coiled coil motif 2. Linker: Sequence without coiled coil prediction. a, d: Key positions in the heptad repeats of MARV NP. Large boxes: conserved amino acids at the coiled coil key positions. Small boxes: Key positions in MARV NP without conservation in Ebola virus NP.
Mentions: We have recently reported that homooligomerization of VP35, the polymerase cofactor, is mediated by a coiled coil motif which leads to the formation of tetramers [10]. Here we show that the predicted coiled coil domains in NP, the major MARV nucleocapsid protein, play an essential role in the formation of NP-NP oligomers and the formation of inclusion bodies which contain preformed NP-induced nucleocapsid-like structures [8]. The nucleoprotein of Ebola virus also contains a region, encompassing amino acids 334 to 363, which has a very high probability to form a coiled coil structure (Fig. 5A). The predicted coiled coil in Ebola virus NP corresponds to the predicted coiled coil C1 of MARV NP. Seven of the nine key residues of C1 are conserved between MARV and ZEBOV (Fig. 5B). Interestingly, the second coiled coil predicted in MARV NP (amino acid positions 371 – 400) is less conserved in Ebola virus NP (Fig. 5B).

Bottom Line: In the present study, a conserved coiled coil motif in the central part of MARV NP was shown to be an important element for the interactions of NP with itself and VP35, the viral polymerase cofactor.Additionally, the coiled coil motif was essential for the formation of NP-induced intracellular inclusions and for the function of NP in the process of transcription and replication of viral RNA in a minigenome system.The coiled coil motif is bipartite, constituted by two coiled coils which are separated by a flexible linker.

View Article: PubMed Central - HTML - PubMed

Affiliation: Philipps-Universität Marburg, Institut für Virologie, Hans Meerwein-Str, 2, 35032 Marburg, Germany. andrea.dicarlo@promega.com

ABSTRACT
The nucleoprotein (NP) of Marburg virus (MARV) is responsible for the encapsidation of viral genomic RNA and the formation of the helical nucleocapsid precursors that accumulate in intracellular inclusions in infected cells. To form the large helical MARV nucleocapsid, NP needs to interact with itself and the viral proteins VP30, VP35 and L, which are also part of the MARV nucleocapsid. In the present study, a conserved coiled coil motif in the central part of MARV NP was shown to be an important element for the interactions of NP with itself and VP35, the viral polymerase cofactor. Additionally, the coiled coil motif was essential for the formation of NP-induced intracellular inclusions and for the function of NP in the process of transcription and replication of viral RNA in a minigenome system. Transfer of the coiled coil motif to a reporter protein was sufficient to mediate interaction of the constructed fusion protein with the N-terminus of NP. The coiled coil motif is bipartite, constituted by two coiled coils which are separated by a flexible linker.

Show MeSH
Related in: MedlinePlus