Differential polyubiquitin recognition by tandem ubiquitin binding domains of Rabex-5

Shin, Dong Hyuk; Lee, Sei Young; Han, Seungsoo; Ren, Shuo; Kim, Soyoun; Aikawa, Yoshikatsu; Lee, Sangho

doi:10.1016/j.bbrc.2012.06.032

Cited by 3 publications

(3 citation statements)

References 27 publications

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“…We found that S5a tUIM that was previously reported to bind K11, K48, and K63 chains also captures M1 polyUb chains (Fig EV1A). The tandem A20_ZnF‐MIU domain of Rabex‐5 previously reported to capture M1‐, K48‐, and K63‐polyUb chains also binds to tetraUb linked via K6, K11, K29, and K33 (Fig EV1A). Our results demonstrate that using a panel of tetraUb chains in Halo‐tagged UBD pull‐down assays can reveal the linkage preference of a given UBD.…”

Section: Resultsmentioning

confidence: 95%

A single MIU motif of MINDY ‐1 recognizes K48‐linked polyubiquitin chains

et al. 2017

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The eight different types of ubiquitin (Ub) chains that can be formed play important roles in diverse cellular processes. Linkageselective recognition of Ub chains by Ub-binding domain (UBD)-containing proteins is central to coupling different Ub signals to specific cellular responses. The motif interacting with ubiquitin (MIU) is a small UBD that has been characterized for its binding to monoUb. The recently discovered deubiquitinase MINDY-1/FAM63A contains a tandem MIU repeat (tMIU) that is highly selective at binding to K48-linked polyUb. We here identify that this linkageselective binding is mediated by a single MIU motif (MIU2) in MINDY-1. The crystal structure of MIU2 in complex with K48-linked polyubiquitin chains reveals that MIU2 on its own binds to all three Ub moieties in an open conformation that can only be accommodated by K48-linked triUb. The weak Ub binder MIU1 increases overall affinity of the tMIU for polyUb chains without affecting its linkage selectivity. Our analyses reveal new concepts for linkage selectivity and polyUb recognition by UBDs.

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Section: Resultsmentioning

confidence: 95%

A single MIU motif of MINDY ‐1 recognizes K48‐linked polyubiquitin chains

et al. 2017

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“…7A and B ). This implied that NS1-70 directly binds to domains A, B, and C of Ku70 with a gradually increased K D value from 0.67 to 0.18 μM, and that the full-length Ku70 has the highest K D value of 0.16 μM, suggesting a high affinity ( 75 – 77 ). We also compared the binding kinetics of purified Ku70, the three Ku70 domains, and GST (as a negative control) together at 2 μM with 2 μM purified NS1-70.…”

Section: Resultsmentioning

confidence: 99%

The Large Nonstructural Protein (NS1) of Human Bocavirus 1 Directly Interacts with Ku70, Which Plays an Important Role in Virus Replication in Human Airway Epithelia

Shao

Ning

Wang

et al. 2022

J Virol

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Human bocavirus 1 (HBoV1), an autonomous human parvovirus, causes acute respiratory tract infections in young children. HBoV1 infects well-differentiated (polarized) human airway epithelium cultured at an air-liquid interface (HAE-ALI). HBoV1 expresses a large nonstructural protein, NS1, that is essential for viral DNA replication. HBoV1 infection of polarized human airway epithelial cells induces a DNA damage response (DDR) that is critical to viral DNA replication involving DNA repair with error-free Y-family DNA polymerases. HBoV1 NS1 or the isoform NS1-70 per se induces a DDR. In this study, using the second-generation proximity-dependent biotin identification (BioID2) approach, we identified that Ku70 is associated with the NS1-BioID2 pulldown complex through a direct interaction with NS1. Bio-layer Interferometry (BLI) assay determined a high binding affinity of the NS1 with Ku70, which has an equilibrium dissociation constant (K D ) value of 0.16 μM and processes the strongest interaction at the C-terminal domain. The association of Ku70 with NS1 was also revealed during HBoV1 infection of HAE-ALI. Knockdown of Ku70 and overexpression of the C-terminal domain of Ku70 significantly decreased HBoV1 replication in HAE-ALI. Thus, our study provides for the first time a direct interaction of a parvovirus large nonstructural protein NS1 with Ku70. IMPORTANCE Parvovirus infection induces a DNA damage response (DDR) that plays a pivotal role in viral DNA replication. The DDR includes activation of ATM (Ataxia telangiectasia mutated), ATR (ATM- and RAD3-related), and DNA-PKcs (DNA-dependent protein kinase catalytic subunit). The large nonstructural protein (NS1) often plays a role in the induction of DDR; however, how the DDR is induced during parvovirus infection or simply by the NS1 is not well studied. Activation of DNA-PKcs has been shown as one of the key DDR pathways in DNA replication of HBoV1. We identified that HBoV1 NS1 directly interacts with Ku70, but not Ku80, of the Ku70/Ku80 heterodimer at a high affinity. This interaction is also important for HBoV1 replication in HAE-ALI. We propose that the interaction of the NS1 with Ku70 recruits the Ku70/Ku80 complex to the viral DNA replication center, which activates DNA-PKcs and facilitates viral DNA replication.

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“…The activation of Rab11a can be triggered via its ubiquitination at K145 by the HECT domain and ankyrin repeat containing E3 ubiquitin protein ligase 1 (HACE1) (Lachance et al, ). The guanine‐nucleotide exchange factors of Rab GTPases, such as Rabex‐5, are also ubiquitinated for recruitment from the cytosol to endosomes (Mattera & Bonifacino, ; Shin et al, ), although the relevant E3 Ub ligases remain to be identified. In addition to the above protein complexes, organization of the cell cytoskeleton is also crucial for vesicular trafficking, and its association with Ub modification is discussed in Section III.3.…”

Section: Ubiquitination In Cellular Events Of Epithelial Morphogenesismentioning

confidence: 99%

Degradation for better survival? Role of ubiquitination in epithelial morphogenesis

Cheng

Zheng

et al. 2018

Biological Reviews

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As a prevalent post-translational modification, ubiquitination is essential for many developmental processes. Once covalently attached to the small and conserved polypeptide ubiquitin (Ub), a substrate protein can be directed to perform specific biological functions via its Ub-modified form. Three sequential catalytic reactions contribute to this process, among which E3 ligases serve to identify target substrates and promote the activated Ub to conjugate to substrate proteins. Ubiquitination has great plasticity, with diverse numbers, topologies and modifications of Ub chains conjugated at different substrate residues adding a layer of complexity that facilitates a huge range of cellular functions. Herein, we highlight key advances in the understanding of ubiquitination in epithelial morphogenesis, with an emphasis on the latest insights into its roles in cellular events involved in polarized epithelial tissue, including cell adhesion, asymmetric localization of polarity determinants and cytoskeletal organization. In addition, the physiological roles of ubiquitination are discussed for typical examples of epithelial morphogenesis, such as lung branching, vascular development and synaptic formation and plasticity. Our increased understanding of ubiquitination in epithelial morphogenesis may provide novel insights into the molecular mechanisms underlying epithelial regeneration and maintenance.

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Differential polyubiquitin recognition by tandem ubiquitin binding domains of Rabex-5

Cited by 3 publications

References 27 publications

A single MIU motif of MINDY ‐1 recognizes K48‐linked polyubiquitin chains

A single MIU motif of MINDY ‐1 recognizes K48‐linked polyubiquitin chains

The Large Nonstructural Protein (NS1) of Human Bocavirus 1 Directly Interacts with Ku70, Which Plays an Important Role in Virus Replication in Human Airway Epithelia

Degradation for better survival? Role of ubiquitination in epithelial morphogenesis

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