NVL2 Is a Nucleolar AAA-ATPase that Interacts with Ribosomal Protein L5 through Its Nucleolar Localization Sequence

Nagahama, Masami; Hara, Yoshimitsu; Seki, Akihiro; Yamazoe, Takeshi; Kawate, Yumiko; Shinohara, Takashi; Hatsuzawa, Kiyotaka; Tani, Kazuhide; Tagaya, Mitsuo

doi:10.1091/mbc.e04-08-0692

Cited by 46 publications

(58 citation statements)

References 37 publications

(40 reference statements)

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“…As has been observed in other proteins targeted to nucleoli (e.g. Nagahama et al, 2004;Horke et al, 2004), the clusters of positively charged amino acids in the jerker peptide likely include signals for nuclear import and nucleolar retention. Even though the jerker espin mRNAs are maintained at wild-type levels, jerker espin proteins do not accumulate in the tissues of jerker mice (Zheng et al, 2000).…”

Section: Discussionmentioning

confidence: 54%

Targeted wild-type and jerker espins reveal a novel, WH2-domain-dependent way to make actin bundles in cells

Loomis

Kelly

Zheng

et al. 2006

Journal of Cell Science

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The espin actin-bundling proteins, which are the target of deafness mutations, are present in the parallel actin bundles of stereocilia and microvilli and appear to increase their steady-state length. Here, we report a new activity of the espins, one that depends on their enigmatic WH2 domain: the ability to assemble a large actin bundle when targeted to a specific subcellular location. This activity was observed for wild-type espins targeted to the centrosome in transfected neuronal cells and for jerker espins targeted to the nucleolus in a wide variety of transfected cells as a result of the frameshifted peptide introduced into the espin C-terminus by the jerker deafness mutation. This activity, which appears specific to espins, requires two espin F-actinbinding sites and the actin-monomer-binding activity of the espin WH2 domain, but can be mimicked by adding a WH2 domain to an unrelated actin-bundling protein, villin. Espins do not activate the Arp2/3 complex in vitro, and bundle assembly is not indicative of in-vitro nucleation activity. Our results suggest a novel way to build actin bundles at specific sites in cells.

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Section: Discussionmentioning

confidence: 54%

Targeted wild-type and jerker espins reveal a novel, WH2-domain-dependent way to make actin bundles in cells

Loomis

Kelly

Zheng

et al. 2006

Journal of Cell Science

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“…How exactly the two NLSs facilitate ORF57 nucleolar localization is unclear. Current thinking states that proteins that localize to the nucleolus do so by means of an interaction with other nucleolar components (3,(26)(27)(28). Therefore, one possible explanation is that either NLS is capable of nuclear import by means of an interaction with importins.…”

Section: Discussionmentioning

confidence: 99%

Nucleolar trafficking is essential for nuclear export of intronless herpesvirus mRNA

Boyne

Whitehouse

2006

Proc. Natl. Acad. Sci. U.S.A.

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The nucleolus is the largest subnuclear structure and is plurifunctional in nature. Here, we demonstrate that nucleolar localization of a key herpesvirus regulatory protein is essential for its role in virus mRNA nuclear export. The herpesvirus saimiri ORF57 protein is a nucleocytoplasmic shuttle protein that is conserved in all herpesviruses and orchestrates the nuclear export of viral intronless mRNAs. We demonstrate that expression of the ORF57 protein induces nucleolar redistribution of human TREX (transcription͞ export) proteins that are involved in mRNA nuclear export. Moreover, we describe a previously unidentified nucleolar localization signal within ORF57 that is composed of two distinct nuclear localization signals. Intriguingly, point mutations that ablate ORF57 nucleolar localization lead to a failure of ORF57-mediated viral mRNA nuclear export. Furthermore, nucleolar retargeting of the ORF57 mutant was achieved by the incorporation of the HIV-1 Rev nucleolar localization signal, and analysis demonstrated that this modification was sufficient to restore viral mRNA nuclear export. This finding represents a unique and fundamental role for the nucleolus in nuclear export of viral mRNA.mRNA export ͉ nucleolus ͉ virus T he eukaryotic cell nucleus is a highly organized environment containing distinct and often dynamic compartments (1). Of these, the nucleolus is the most prominent, and, for many years, its exclusive role was thought to be the site of ribosomal RNA transcription, processing, and assembly into the ribosome subunits (2). Recent studies, however, suggest that it has additional nonclassical roles in many aspects of cell biology, including cell cycle regulation, viral replication, tumorigenesis, and cellular stress responses (3-5). This plurifunctional nature of the nucleolus has been highlighted by extensive proteomic analysis of human nucleoli (6, 7). To date, the nucleolar proteome database archives 728 nucleolar proteins, and functional classification of these proteins reinforces the multiple roles of the nucleolus (8). Furthermore, there is constant dynamic trafficking of nucleolar proteins, and this concomitant dynamic nature of nucleolar structure may provide regulation of nonclassical nucleolar functions.Interestingly, an increasing number of key proteins from both RNA and DNA viruses have been shown to localize to the nucleolus. These proteins include those encoded by viruses such as coronaviruses, influenza, HIV-1, adenoviruses, and herpesviruses (9). Therefore, virus-nucleolar interactions are likely to have important implications in the life cycle of many viruses. However, at present, the precise functional role of these virus-nucleolar colocalizations has not been determined. One such key viral protein that traffics to the nucleolus is the herpesvirus saimiri (HVS) ORF57 protein. HVS is the prototype ␥-2 herpesvirus, or rhadinovirus (10), which has become an important family of viruses since the identification of the first human ␥-2 herpesvirus, the oncogenic Kaposi's sarcoma-associ...

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“…NVL2 resides in the nucleolus, where it is involved in the biogenesis of the 60 S ribosomal subunit. In this pathway, the ribosomal subunit RPL5 and the DNA helicase DOB1 have been identified as NVL2-binding proteins (6,7). Other well known NVL orthologs include smallminded (smid), a Drosophila ortholog (7,8); a CED-4-interacting protein, Mac-1, a Caenorhabditis elegans ortholog (9); and a more distant Saccharomyces cerevisiae homolog, Rix7p (10,11).…”

Section: Nuclear Valosin-containing Protein (Vcp)mentioning

confidence: 99%

“…The two proteins reside in different subcellular locations; VCP/p97 is in the cytoplasm, nucleus, and endoplasmic reticulum membrane, whereas NVL is exclusively localized in the nucleolus (5,6). The amino acid sequences of the N-terminal regions are variable, in contrast to the high sequence conservation of the AAA domains.…”

Section: Nuclear Valosin-containing Protein (Vcp)mentioning

confidence: 99%

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Structure and Function of the N-terminal Nucleolin Binding Domain of Nuclear Valosin-containing Protein-like 2 (NVL2) Harboring a Nucleolar Localization Signal

Fujiwara

Goda

et al. 2011

Journal of Biological Chemistry

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The N-terminal regions of AAA-ATPases (ATPase associated with various cellular activities) often contain a domain that defines the distinct functions of the enzymes, such as substrate specificity and subcellular localization. As described herein, we have determined the solution structure of an N-terminal unique domain isolated from nuclear valosin-containing protein (VCP)-like protein 2 (NVL2 UD ). NVL2 UD contains three ␣ helices with an organization resembling that of a winged helix motif, whereas a pair of ␤-strands is missing. The structure is unique and distinct from those of other known type II AAA-ATPases, such as VCP. Consequently, we identified nucleolin from a HeLa cell extract as a binding partner of this domain. Nucleolin contains a long (ϳ300 amino acids) intrinsically unstructured region, followed by the four tandem RNA recognition motifs and the C-terminal glycine/arginine-rich domain. Binding analyses revealed that NVL2 UD potentially binds to any of the combinations of two successive RNA binding domains in the presence of RNA. Furthermore, NVL2 UD has a characteristic loop, in which the key basic residues RRKR are exposed to the solvent at the edge of the molecule. The mutation study showed that these residues are necessary and sufficient for nucleolin-RNA complex binding as well as nucleolar localization. Based on the observations presented above, we propose that NVL2 serves as an unfoldase for the nucleolin-RNA complex. As inferred from its RNA dependence and its ATPase activity, NVL2 might facilitate the dissociation and recycling of nucleolin, thereby promoting efficient ribosome biogenesis.

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NVL2 Is a Nucleolar AAA-ATPase that Interacts with Ribosomal Protein L5 through Its Nucleolar Localization Sequence

Cited by 46 publications

References 37 publications

Targeted wild-type and jerker espins reveal a novel, WH2-domain-dependent way to make actin bundles in cells

Targeted wild-type and jerker espins reveal a novel, WH2-domain-dependent way to make actin bundles in cells

Nucleolar trafficking is essential for nuclear export of intronless herpesvirus mRNA

Structure and Function of the N-terminal Nucleolin Binding Domain of Nuclear Valosin-containing Protein-like 2 (NVL2) Harboring a Nucleolar Localization Signal

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