Using sorting signals to retain proteins in endoplasmic reticulum

Pelham, Hugh R.B.

doi:10.1016/s0076-6879(00)27283-2

Cited by 18 publications

(14 citation statements)

References 21 publications

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“…It is important to generate (Man) 5 -(GlcNAc) 2 structures in vivo, and early in the secretory pathway, if subsequent conversion to complex glycans is to be achieved. Although it is well established that the ERD2-based retrieval system is leaky and retention of the HDEL-tagged mannosidase in the secretory pathway cannot be ensured (26), the same concern is emphasized further by the striking difference demonstrated here between the two different C. elegans ␣-1,2-mannosidase IB fusion constructs. Thus, the use of a series of different fusion constructs with many differentially localized type II domains has allowed us to screen for chimera with activity that is completely in vivo.…”

Section: Discussionmentioning

confidence: 85%

Use of combinatorial genetic libraries to humanize N-linked glycosylation in the yeast Pichia pastoris

Choi

Bobrowicz²,

Davidson³

et al. 2003

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

325

211

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The secretory pathway of Pichia pastoris was genetically re-engineered to perform sequential glycosylation reactions that mimic early processing of N-glycans in humans and other higher mammals. After eliminating nonhuman glycosylation by deleting the initiating α-1,6-mannosyltransferase gene from P. pastoris, several combinatorial genetic libraries were constructed to localize active α-1,2-mannosidase and human β-1,2-N-acetylglucosaminyltransferase I (GnTI) in the secretory pathway. First, >32 N-terminal leader sequences of fungal type II membrane proteins were cloned to generate a leader library. Two additional libraries encoding catalytic domains of α-1,2-mannosidases and GnTI from mammals, insects, amphibians, worms, and fungi were cloned to generate catalytic domain libraries. In-frame fusions of the respective leader and catalytic domain libraries resulted in several hundred chimeric fusions of fungal targeting domains and catalytic domains. Although the majority of strains transformed with the mannosidase/leader library displayed only modest in vivo [i.e., low levels of mannose (Man)5-(GlcNAc)2] activity, we were able to isolate several yeast strains that produce almost homogenous N-glycans of the (Man)5-(GlcNAc)2 type. Transformation of these strains with a UDP-GlcNAc transporter and screening of a GnTI leader fusion library allowed for the isolation of strains that produce GlcNAc-(Man)5-(GlcNAc)2 in high yield. Recombinant expression of a human reporter protein in these engineered strains led to the formation of a glycoprotein with GlcNAc-(Man)5-(GlcNAc)2 as the primary N-glycan. Here we report a yeast able to synthesize hybrid glycans in high yield and open the door for engineering yeast to perform complex human-like glycosylation

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Section: Discussionmentioning

confidence: 85%

Use of combinatorial genetic libraries to humanize N-linked glycosylation in the yeast Pichia pastoris

Choi

Bobrowicz²,

Davidson³

et al. 2003

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

325

211

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“…In addition to KDEL and KKXX (Lys-Lys-X-X), the wellknown ER retention signals in most luminal ER proteins (17)(18)(19), it is widely accepted that RXR is the minimal motif of 20). We found that ERIS contains a RYR at position 76-78aa (between TM2 and TM3) and an RIR at position 178-180aa (right after TM4) (Fig.…”

Section: Eris Localizes To Er Membranementioning

confidence: 64%

ERIS, an endoplasmic reticulum IFN stimulator, activates innate immune signaling through dimerization

Sun

Chen

et al. 2009

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

714

625

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We report here the identification and characterization of a protein, ERIS, an endoplasmic reticulum (ER) IFN stimulator, which is a strong type I IFN stimulator and plays a pivotal role in response to both non-self-cytosolic RNA and dsDNA. ERIS (also known as STING or MITA) resided exclusively on ER membrane. The ER retention/ retrieval sequence RIR was found to be critical to retain the protein on ER membrane and to maintain its integrity. ERIS was dimerized on innate immune challenges. Coumermycin-induced ERIS dimerization led to strong and fast IFN induction, suggesting that dimerization of ERIS was critical for self-activation and subsequent downstream signaling.innate immunity ͉ type I IFN ͉ functional cDNA library screening ͉ cytosolic RNA and dsDNA ͉ ER retention signal M icrobial infection-induced host immune responses are initiated by the germline-encoded pattern recognition receptors, which recognize components specific to microorganisms. There are 3 major classes of such receptors: Toll-like receptors (TLRs), RIG-I-like helicases (RLHs) and NOD-like receptors (1). During infection, nucleic acids derived from microbes are recognized by TLRs and RLHs, which then trigger a series of signaling events leading to the production of type I IFNs and proinflammatory cytokines.RLHs have recently been identified to sense the invading viruses in the cytoplasm. Unlike TLRs, which are expressed in specific cells like macrophages and dendritic cells, RLHs are found in most cell types (2). They contain caspase recruitment domain (CARD) and DExD/H helicase domain. RLHs interact with microbial nucleotides through their helicase domain. The N-terminal CARDs are responsible for activating downstream signaling pathways that mediate type I IFN production. Genetic analyses demonstrate that RIG-I and MDA5 sense distinct types of viruses (3-5). RIG-I and MDA5 use a common adaptor molecule, IPS-1 (also known as Cardif, MAVS, or VISA) (6-9). IPS-1 is found to reside on the mitochondrial membrane by its C-terminal transmembrane (TM) domain. It also contains a CARD-like domain at its N-terminus, which mediates the interaction with MDA5 or RIG-I. IPS-1 transmits the signal to TANK-binding kinase-1 (TBK1)/I B kinase i (IKKi; also known as IKK ) and the IKK complex to activate interferon regulatory factor (IRF)-3/IRF-7 and NF-B, respectively, collectively eliciting innate antiviral immune responses, including type I IFN production.On the other hand, dsDNA in the cytosol, for example, genomic DNA from intracellular bacteria (e.g., Listeria, Legionella), also causes a strong host immune response independent of TLRs, leading to the induction of type I IFN. A recent report has indicated that the molecule DAI (also known as ZBP1) might serve as a cytosolic dsDNA sensor (10). However, ZBP1 Ϫ/Ϫ cells showed normal type I IFN production in response to dsDNA stimulation (11). Meanwhile, reports showed that IPS-1/Cardif/MAVS/VISA was not required for dsDNA-caused innate immune activation (12).The signaling induced by cytoplasmic dsDNA leading ...

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“…1). KDEL (Lys-AspGlu-Leu,), KKXX (Lys-Lys-X-X; X represents any resides), and RXR (Arg-X-Arg; X represents any resides) are well-known ER retention signals in most luminal ER proteins (34)(35)(36). The RXR motif is widely accepted as the minimal motif for retention/retrieval of resident ER proteins (36).…”

Section: Discussionmentioning

confidence: 99%

Chicken STING Mediates Activation of the IFN Gene Independently of the RIG-I Gene

Cheng

Sun

Wang

et al. 2015

The Journal of Immunology

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Stimulator of IFN genes (STING) is an adaptor that functions downstream of retinoic acid–inducible gene I (RIG-I) in mammalian cells; however, RIG-I is absent in chickens. We identified chicken STING (chSTING) as a critical mediator of virus-triggered type I IFN signaling in RIG-I–null chicken cells. Overexpression of chSTING in DF-1 cells inhibited Newcastle disease virus and avian influenza virus (AIV) viral replication and activated IRF-7 and NF-κB to induce expression of type I IFNs. Knockdown of endogenous chSTING abolished virus-triggered activation of IRF-7 and IFN-β and increased viral yield. chSTING was a critical component in the virus-triggered IRF-7 activation pathway and the cellular antiviral response. chSTING predominantly localized to the outer membrane of the endoplasmic reticulum and was also found in the mitochondrial membrane. Furthermore, knockdown of chSTING blocked polyinosinic-polycytidylic acid–, poly(deoxyadenylic-deoxythymidylic) acid–, and melanoma differentiation–associated gene 5 (MDA5)-stimulated induction of IFN-β. Coimmunoprecipitation experiments indicated that chicken MDA5 could interact with chSTING, and this interaction was enhanced by ectopically expressed chicken mitochondrial antiviral-signaling protein. Together, these results indicated that chSTING is an important regulator of chicken innate immune signaling and might be involved in the MDA5 signaling pathway in chicken cells. These results help with understanding the biological role of STING in innate immunity during evolution.

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Using sorting signals to retain proteins in endoplasmic reticulum

Cited by 18 publications

References 21 publications

Use of combinatorial genetic libraries to humanize N-linked glycosylation in the yeast Pichia pastoris

Use of combinatorial genetic libraries to humanize N-linked glycosylation in the yeast Pichia pastoris

ERIS, an endoplasmic reticulum IFN stimulator, activates innate immune signaling through dimerization

Chicken STING Mediates Activation of the IFN Gene Independently of the RIG-I Gene

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