476

ER-associated degradation in protein quality control and

cellular regulation
Randolph Y Hampton
The ER-associated degradation (ERAD) pathway directs p53 in both health and disease. In these and innumerable
ubiquitin-mediated degradation of a variety of ER-associated other cases, features that uniquely specify the targeted
misfolded and normal proteins. Recent studies have delineated protein are recognised, usually by ubiquitin ligases, to
the molecular machinery responsible for protein ubiquitination effect selective degradation.
and highlighted mechanistic questions surrounding the
recognition, extraction and proteasomal destruction of the By contrast, quality-control degradation requires recognition
diverse array of ERAD substrates. Consideration of separate of hallmarks of misfolding shared by a diverse group of
lines of work on this versatile pathway now indicate that proteins with no sequence similarity. Such criteria might
despite its central role as an avenue of cellular quality control, seem less useful for cellular regulation of functional
ERAD is also harnessed for feedback regulation of sterol cellular proteins. Nevertheless, studies indicate that the
synthesis, and most likely numerous other cellular processes. essential enzyme 3-hydroxy 3-methylglutaryl coenzyme A
These studies give ERAD a larger role in cellular function, and reductase (HMG-CoA reductase, HMGR) can undergo
imply that cellular quality-control pathways could be widely physiologically regulated degradation by ERAD. Beyond
employed in both natural and pharmaceutical control of this demonstrated role in sterol regulation, regulation by
individual proteins. quality-control degradation is likely to be used in many
cellular situations, and may present new avenues for
Addresses pharmaceutical modulation of protein levels. Specifically,
Section of Cell and Developmental Biology, UCSD Division of Biology I will discuss new developments in the understanding of
#0347, Room 2100E, Pacific Hall, La Jolla, CA 92093-0347, USA; ERAD, including work on specific ubiquitin ligases,
e-mail: rhampton@ucsd.edu
substrate recognition, retrotranslocation, and the role of
Current Opinion in Cell Biology 2002, 14:476482 this broadly used pathway in the control of sterol synthesis.
0955-0674/02/$ see front matter
2002 Elsevier Science Ltd. All rights reserved. HMG-CoA reductase: a regulated ER
degradation substrate
Abbreviations HMGR is a rate-limiting enzyme of the mevalonate path-
CFTR cystic fibrosis transmembrane conductance regulator
CPY carboxypeptidase Y
way, by which sterols and a variety of essential isoprenoids
DER degradation in the ER are made. In eukaryotic cells, HMGR is controlled, in
ERAD ER-associated degradation part, by regulated ER degradation [58]. When flux
FPP farnesyl pyrophosphate through the sterol pathway is high, degradation rate is
HMGR 3-hydroxy 3-methylglutaryl-CoA reductase
HRD HMG-CoA reductase degradation
high and levels of the protein tend to be low. When flux is
SCAP SREBP cleavage-activating protein low, degradation rate is low and enzyme levels tend to
UBC ubiquitin-conjugating enzyme increase. Depending on cell type and signal strength,
UPR unfolded protein response HMGR half life can vary between >10 h, and <20 min.
HMGR is an ER-resident membrane protein, with a
Introduction: degradation for quality control highly conserved, catalytic carboxyl terminus attached to
or regulation a polytopic amino-terminal transmembrane anchor. The
Recently, there has been an explosion of interest and infor- transmembrane region is essential for feedback control of
mation about ER-associated protein degradation (ERAD) stability and will impart regulated degradation to fusion
[13]. This degradation process is responsible for the proteins without catalytic activity.
destruction of both integral membrane and lumenal
proteins, and functions in protein quality control, where So, what is the mechanism of regulated HMGR destruc-
damaged or unfolded proteins are selectively targeted for tion, and what is the connection to the ERAD pathway?
degradation, while correctly folded ones are spared.
Yeast HMG2 degradation: the HRD pathway
Both normal and misfolded proteins undergo highly and ER ubiquitination
specific degradation. Selective degradation of correctly The yeast HMGR isozyme Hmg2p undergoes regulated
folded proteins underlies cellular regulation of many degradation in a manner very similar to the mammalian
processes [4]. Examples include degradation of cyclins or enzyme. Isolation of HRD (HMG-CoA reductase
their inhibitors to control the cell cycle, destruction degradation) genes required for HMGR degradation
of HIF in response to changing O2 levels, degradation of has revealed proteins involved all along the degrad-
IB to launch innate immunity responses, light-induced ation pathway ([9,10,11]; N Bays and R Hampton,
proteolysis of phytochrome, or regulated degradation of unpublished data).
 ER-associated degradation in protein quality control and cellular regulation Hampton 477

Figure 1

Two ER-associated ubiquitin ligases. The

Hrd1p/Hrd3p complex is shown on the left,
and the Doa10p ligase is shown on the right. Ubc7p Ubc7p Ubc6p
The functionally associated ubiquitin

Cue1p

Cue1p
RING Ubc1p RING
conjugating enzymes (E2s) are also shown,
along with Cue1p, which anchors Ubc7p to N
the lumenal face of the ER membrane. In both Cytosol C
cases, a multi-spanning RING-domain protein
(Hrd1p or Doa10p) is a key component of ER membrane
the complex. In all likelihood, still other Lumen
ubiquitin E3s are involved in ER degradation Hrd1p/Der3p
of some substrates. Hrd3p Doa10p

Current Opinion in Cell Biology

HRD2 encodes the RPN1 subunit of the 26S proteasome, of CPY* and regulated degradation of Hmg2p, the same
and other HRD mutants are alleles of numerous proteasomal machinery is employed for ubiquitination. The encoded
genes. Many proteins are targeted to the proteasome by DER proteins include Der3p, which is identical to Hrd1p
covalent addition of multiple copies of the 7.6 kDa protein [21], Hrd3p [22], the E2s Ubc7p (Der2p [19]) and Ubc1p
tag ubiquitin to form a multiubiquitin chain [4]. Specific [23], as well as several proteins specifically required only
ubiquitination is catalyzed by a ubiquitin protein ligase, or for lumenal substrates (e.g. Der1p). Work by numerous
E3, that brokers the transfer of ubiquitin from a ubiquitin- laboratories has shown that this pathway degrades a variety
conjugating enzyme (UBC, E2) to the target protein or the of misfolded or unassembled proteins with no similarity in
growing multiubiquitin chain. Hrd1p and Hrd3p are key sequence [24,25,26].
components of an ER-anchored ubiquitin ligase of the
growing RING-motif family of E3s. The cytosolic RING- These studies using model substrates imply that the
H2 domain of Hrd1p is required for Hmg2p ubiquitination HRD/DER pathway is used for degradation of naturally
in vivo, and displays ligase activity in vitro [10]. Hrd3p arising misfolded proteins. This idea is supported by the
functions in the ER lumen in multiple aspects of Hmg2p observations that hrd1- or hrd3-null mutants are more
degradation, including transmembrane regulation of sensitive to applied ER stresses, and have higher levels of
Hrd1p self-degradation [12]. Hrd1p and Hrd3p form a unfolded ER proteins in the absence of stress, as measured
complex that mediates interaction of E2 with degradation using reporters regulated by the unfolded protein response
substrate [13]. The HRD ligase uses the ER-anchored (UPR), a signaling pathway that is keyed to levels of lumenal
Ubc7p (Hrd10p) as the principle UBC, and, to a lesser misfolded proteins [23,27]. This latter result is particu-
extent, Ubc1p [10]. Ubc7p is anchored to the ER by larly intriguing because it implies that unfolded protein
Cue1p [2] (Hrd13p). Figure 1 depicts the HRD ligase with levels in the ER are kept in check by continuous degradation
relevant E2s. The ubiquitinproteasome pathway is also of misfolded proteins either nascent ones that partition
responsible for HMGR degradation in mammals [14,15]. into the degradation pathway while still unfolded, or those
Although the machinery of mammalian HMGR ubiquiti- that become misfolded for other reasons.
nation and membrane extraction has not been determined,
there are mammalian homologues of most HRD compo- More recent work has led to the realisation that ERAD is
nents that are good candidates for the job in larger complex and multifaceted. Below, I discuss and delineate
eukaryotes [1618]. some of the new findings of this burgeoning field, including
the unexpected use of ERAD for the regulation of HMGR
CPY* degradation and the DER pathway of and probably other proteins.
ER-associated degradation
The genetics of ER quality-control degradation has been Between ubiquitination and the proteasome:
analysed by the Wolf laboratory. Their studies focused on Cdc48 and retrotranslocation
CPY*, a mutant version of the normally vacuole-bound Removal of both membrane proteins and purely lumenal
carboxypeptidase Y that is retained in the lumen of the ERAD substrates presents a significant energy barrier. A
ER, where it is degraded with a half life of around 20 minutes. variety of current studies focus on a molecular complex
Genes for CPY* degradation in the ER (DER) have been that appears to manage this problem. The more recently
identified, revealing the unanticipated role of the cytosolic characterized Hrd4p functions after Hmg2p ubiquitina-
ubiquitinproteasome pathway for the degradation of this tion, promoting delivery of the ubiquitinated Hmg2p to
demonstrably lumenal substrate [19,20]. Despite the the proteasome [11]. Hrd4p is identical to Npl4p, and
conceptual disparity between quality control destruction exists in a complex with AAA-ATPase Cdc48p and Ufd1p
478 Membranes and organelles

to promote proteasomal degradation or processing of a chaperones (i.e. substrate recognition) may exist in ERAD
growing number of ubiquitinated substrates in mammals ligases as well. It is interesting that the Hrd3p lumenal
and yeast [11,2833]. The Cdc48pNpl4pUfd1p domain has numerous tetratricopeptide repeats, indepen-
complex has been postulated to effect removal of ubiquiti- dently implicated in interactions between chaperones and
nated proteins from the ER and/or multiprotein complexes various binding partners [45]. Conversely, it may be that
(see [34] for a more detailed review). the ligase subunits themselves participate in recognition of
misfolded proteins. The Hrd1p cytosolic RING domain
The ER translocation channel Sec61p has also been has the intrinsic capacity to ubiquitinate a model misfolded
proposed to be involved in retrotranslocation of ERAD protein in vitro [10], a behaviour that might reflect an
substrates from the ER compartment, providing a route for aspect of chaperone-independent identification of quality-
peptide exit [3537]. This might be fuelled by Cdc48p control substrates.
ATPase activity. Although evidence for the role of Sec61p is
not definitive, this reasonable possibility will certainly be Is Golgi function required in ER-associated
an ongoing focus that will lead to more mechanistic resolution. degradation of some proteins?
Several laboratories have made the surprising discovery
Other ER ubiquitin ligases: a growing group that the route of ER degradation taken by soluble lumenal
of guardians proteins and ER-membrane-anchored proteins may be
The HRD/DER complex and its ancillary proteins can different. Experiments using mutants deficient in secretory
degrade an impressive variety of substrates. However, there pathway traffic [26,46], and direct biochemical studies
are numerous misfolded proteins that are ubiquitinated using in vitro budding assays [26], indicate that lumenal
by the ER-associated E2 Ubc7p in a Hrd1p-independent proteins may require a visit to the Golgi compartment
manner. These non-Hrd1p substrates include mutant before their destruction by ERAD. These authors propose
ER-retained uracil permease [25], unassembled Vph1p that lumenal proteins must be cycled through the Golgi
[38], and mammalian CFTR (cystic fibrosis transmembrane compartment, and then retrieved to the ER before being
conductance regulator) expressed in yeast [39]. competent for degradation. By contrast, integral membrane
ERAD substrates do not generally have this requirement,
These exceptions indicate that other ubiquitin ligases although not enough substrates have been tested to
participate in ERAD. In fact recently, a new yeast ER- strengthen this generalization. Genes required for aspects
resident multi-spanning ubiquitin ligase, Doa10p, has been of this process BST1 [26], ERV29 and ERV14 [46]
cloned and characterized [40]. Like Hrd1p, Doa10p has a have been implicated previously in aspects of ERGolgi
role in ERAD, as indicated by increased UPR signaling in transport, although BST1 appears to be involved specifically
doa10-nulls. In contrast to Hrd1p, Doa10 uses E2s Ubc7p in transport of misfolded proteins, adding another process
and Ubc6p, and its model substrates are Hrd1p-indepen- where recognition of features of misfolding occurs.
dent (Figure 1). Doa10p and Hrd1p appear to work
together to maintain acceptable levels of misfolded ER pro- At present, it is not clear what features of a Golgi visit
teins: loss of both proteins causes a larger UPR than loss of would render substrates ERAD-competent. One possibility
either alone. Interestingly, the ER degradation of Ole1p has is that some sort of glycosylation or processing of carbohy-
been reported to be unaffected by loss of either Hrd1p or drates must occur. Alternatively, it could be that other
Doa10p [28], implying that other ER E3s await discovery. Golgi modifications are involved, or that the retrotranslocation
process itself allows efficient presentation of lumenal substrates
How are misfolded proteins recognised for to the ERAD machinery. However, another interpretation
quality control degradation? cannot be ignored [47]. Mutants deficient in ER-to-Golgi
The mechanism of misfolded protein recognition in cycling will most certainly have effects on the ER
ERAD remains an open question. It is not surprising that compartment, and far more work must be done to decide
chaperones have been implicated in several studies. It if these observations are reflecting a deficiency in ER
appears that the lumenal heat shock protein 70 (Hsp70) function or a bone fide requirement for Golgi visitation for
Kar2p may be more important in degradation of lumenal lumenal ERAD. Whatever the mechanism, these new
ERAD substrates [35], whereas the four cytosolic Hsp70s, observations make ER degradation a far more dynamic
Ssa14, play a more significant role in degradation of process, allowing for a whole new layer of stability regula-
integral membrane substrates [39]. Only a small number tion based on altering ER exit or return.
of substrates have been examined so far, however.
Regulation of Hmg2p: HRDing a normal
In recent studies, a chaperone-containing ubiquitin ligase protein into the not-OK corral
was described. It is formed by mutual binding of the U-box The ongoing body of work on many aspects of ERAD
protein CHIP with a chaperone and an E2, leading to a shows that unfolded proteins can undergo constitutive
complex that specifically ubiquitinates chaperone- degradation by a variety of mechanisms. However, unlike
recognised misfolded proteins [4143], including the any of the typical ERAD substrates, HRD-dependent
ER membrane protein CFTR [44]. A similar role for degradation of Hmg2p is subject to physiological feedback
 ER-associated degradation in protein quality control and cellular regulation Hampton 479

control. This exception is interesting in its own right, by Figure 2

virtue of the connection to cholesterol, but also indicates
the strong possibility that ER quality-control pathways are
involved in unknown aspects of cellular regulation. Hmg2p Hmg2p

Hmg2p ubiquitination is positively regulated by the

15-carbon mevalonate pathway intermediate farnesyl High FPP
pyrophosphate (FPP), or a molecule derived from it [48].
Altering cellular FPP levels causes parallel alterations in Low FPP
Hmg2p degradation rate, and this regulation requires
Cod1p/Spf1p, an ER-localized P-type ATPase [49,50].
ERAD
Maximal response to FPP requires an oxysterol in yeast as
Current Opinion in Cell Biology
well [51]. Similarly, mammalian HMGR degradation is
keyed to levels of an FPP-derived signal that works in con-
The structural transition model for regulated degradation of Hmg2p. In
cert with sterols [52,53]. Regulated degradation of Hmg2p
cells with high levels of FPP, Hmg2p is recognised as an ERAD
is a unique property of the protein, and is highly sensitive quality-control substrate by the HRD/DER machinery. Lowering FPP
to alterations in sequence [54]. Other HRD substrates, levels or glycerol cause a rapid, reversible stabilization of Hmg2p.
including damaged versions of Hmg2p, undergo constitu- Although the structural change is depicted as graphically dramatic, it
may be that subtle determinants are presented or unveiled when FPP
tive, unregulated degradation. The entire pool of Hmg2p
is high, triggering recognition as an ERAD substrate. Regulated
is subject to regulated entry into the HRD pathway, and transitions to ERAD or other quality-control substrates have broad
mature protein can be rapidly mobilized to enter the HRD potential as both undiscovered mechanisms in normal cellular
pathway by elevating FPP [8]. regulation and as undeveloped axes for chemical modulation of
individual proteins in vivo.
The structural transition model for
regulated ERAD
How is Hmg2p allowed regulated entry into a quality- to that caused by lowering FPP-derived signals? How does
control pathway? The most direct model is that Hmg2p FPP or its derivatives cause this change? Limited proteolysis
can adopt or reveal the structural features of a quality- studies on microsomal Hmg2p appear to be a promising
control substrate when degradation signals are high avenue [13], and are being refined to best address this
(Figure 2). This idea was tested by adapting an approach structural transition hypothesis.
used in the study of CFTR F508, the major disease-
causing form of the cystic fibrosis protein CFTR. CFTR This model of yeast Hmg2p regulation has parallels to
F508 is functional, but folds so slowly that it is degraded mammalian HMGR. It has been suggested that mammalian
by ubiquitin-mediated ER degradation before it can attain HMGR is susceptible to ERAD when monomeric, is stable
a degree of folding maturity sufficient to exit the ER and when a dimer, and that this transition is physiologically
move to the plasma membrane. The folding defect of regulated [58]. This can be viewed as regulated transition to
CFTR F508, and the resulting lack of cell surface a quality control structure, because many quality control
chloride ion conductance, can be alleviated in living cells substrates are unassembled (monomeric) subunits of
by treatment with the chemical chaperone glycerol [55,56]. multimeric structures. In fact, it could be that the same
This class of structurally diverse compounds promotes the occurrence underlies the HRD-sensitive state of Hmg2p.
folding of proteins in vitro and in vivo [57].
Biological and medical implications of
If Hmg2p is recognised as a quality-control substrate, then regulation by quality control
glycerol might be expected to similarly stabilize this The programmed entry of Hmg2p into the ER quality-
protein in living yeast cells. In conditions where signals for control pathway is an example of a mode of protein
degradation are high, glycerol and other chemical chaper- regulation that could occur in many circumstances.
ones cause rapid (5 min) stabilization of Hmg2p or the Considering the number of separate routes by which
regulated Hmg2pGFP reporter [13]. The extent of sta- ERAD appears to occur, it is quite possible that other
bilization caused by glycerol is identical to that observed proteins are similarly regulated. It has been reported that
when physiological signals for degradation are lowered yeast 9 fatty acid desaturase, or Ole1p, undergoes regulated
with drugs, and the maximal effects of glycerol and physi- degradation in the ER membrane, and is thus a reasonable
ological stabilization are not additive. Glycerol treatment candidate for regulated ERAD [28].
affects neither the HRD machinery nor the mevalonate
pathway. These simple studies engender several more The idea of regulation by ER quality-control mechanisms
refined questions. For example, are there directly observable can transcend degradation. Mammalian SREBP cleavage-
changes in Hmg2p structure that can be observed by activating protein (SCAP), which shares with both HMGR
manipulating the signaling pathway, or by applying chemical and, to a lesser extent, Hmg2p a transmembrane motif
chaperones? Is the stabilized state caused by glycerol similar known as the sterol-sensing domain, is retained in the
480 Membranes and organelles

mammalian ER when sterol levels are elevated [59]. This 6. Chun KT, Simoni RD: The role of the membrane domain in the
regulated degradation of 3-hydroxy-3-methylglutaryl coenzyme A
sterol-triggered retention is central to SCAP-mediated reductase. J Biol Chem 1992, 267:4236-4246.
regulation of the SREBP transcriptional regulator. 7. Hampton RY, Rine J: Regulated degradation of HMG-CoA
Although degradation is not the final response, ER retention reductase, an integral membrane protein of the endoplasmic
is another frequently described cellular response to misfolded reticulum, in yeast. J Cell Biol 1994, 125:299-312.
proteins. Perhaps SCAP undergoes a sterol-mediated 8. Hampton RY, Bhakta H: Ubiquitin-mediated regulation of
3-hydroxy-3-methylglutaryl-CoA reductase. Proc Natl Acad Sci
structural transition that renders it susceptible to retention USA 1997, 94:12944-12948.
by ER quality-control mechanisms?
9. Hampton RY, Gardner RG, Rine J: Role of 26S proteasome and
HRD genes in the degradation of 3-hydroxy 3-methylglutaryl-CoA
Cellular use of quality-control pathways need not be reductase, an integral endoplasmic reticulum membrane protein.
Mol Biol Cell 1996, 7:2029-2044.
restricted to the ER. Selective destruction of misfolded
proteins occurs in numerous eukaryotic cellular compart- 10. Bays NW, Gardner RG, Seelig LP, Joazeiro CA, Hampton RY:
Hrd1p/Der3p is a membrane-anchored ubiquitin ligase required
ments, and in prokaryotes, and each specific pathway has for ER-associated degradation. Nat Cell Biol 2001, 3:24-29.
the potential for regulatory employment. This and Gardner et al. (2000) [12] demonstrate that Hrd1p is the rate-
limiting ubiquitin ligase for degradation of Hmg2p (and other ER proteins) in
yeast, and that Hrd1p, with Hrd3p, forms a transmembrane ubiquitin ligase
The example set by Hmg2p also suggests a general strategy that functions on both sides of the membrane. See also Kumagai et al.
(1995) [5] and Gardner et al. (2001) [13].
for controlling proteins by pharmaceutical means. In the
11. Bays NW, Wilhovsky SK, Goradia A, Hodgkiss-Harlow K,
broadest sense, a molecular signal causes HMGR to Hampton RY: HRD4/NPL4 is required for the proteasomal
become susceptible to quality-control degradation. processing of ubiquitinated ER proteins. Mol Biol Cell 2001,
Perhaps it is possible to discover small molecules that, 12:4114-4128.
This, together with [2833], all released in a few months of each other,
upon specifically binding to a target protein, similarly place a new component, the Cdc48 AAA ATPase and its complex partners,
cause that protein to present or acquire features of a in the middle of the ER degradation pathway, between ubiquitination and
proteasomal degradation or processes [31,33], both in yeast and in
quality-control substrate and undergo selective, rapid mammals [30].
degradation. Considering the ubiquity of quality-control 12. Gardner RG, Swarbrick GM, Bays NW, Cronin SR, Wilhovsky S,
pathways, this highly specific strategy of protein targeting Seelig L, Kim C, Hampton RY: Endoplasmic reticulum degradation
could be very generally effective if systematic ways to requires lumen to cytosol signaling. Transmembrane control of
Hrd1p by Hrd3p. J Cell Biol 2000, 151:69-82.
discover and develop such protein quality antagonists can See annotation Bays et al. (2001) [10].
be devised. However, working out the detailed mechanisms 13. Gardner RG, Shearer AG, Hampton RY: In vivo action of the HRD
involved in recognition and destruction of quality-control ubiquitin ligase complex: mechanisms of endoplasmic reticulum
quality control and sterol regulation. Mol Cell Biol 2001,
protein is a sine qua non for the eventual realisation of this 21:4276-4291.
novel strategy. This study demonstrates that glycerol stabilizes Hmg2p but does not affect
the HRD pathway nor the generation of signals that programme Hmg2p
degradation. Thus, the Hmg2p molecule behaves like a misfolded protein in
Acknowledgements cellular conditions where degradation signals are elevated.
I thank members of the Hampton lab for stimulating discussions and
maintenance of the data stream. I thank Mark Hochstrasser, Davis Ng, 14. McGee TP, Cheng HH, Kumagai H, Omura S, Simoni RD:
Jeff Brodsky, and Dieter Wolf for help with ERAD-related issues, along Degradation of 3-hydroxy 3-methylglutaryl-CoA reductase in
with David Katzman, Kaustuv Roy, David Rudner, Irene Folk and endoplasmic reticulum membranes is accelerated as a result of
increased susceptibility to proteolysis. J Biol Chem 1996,
Vivek Malhotra for help with numerous other issues. Omissions of relevant
271:25630-25638.
work were not intentional acts of isolation, but rather a side-effect of
imposed spatial restrictions. This work was supported by a grant from the 15. Ravid T, Doolman R, Avner R, Harats D, Roitelman J: The ubiquitin
National Institutes of Health. proteasome pathway mediates the regulated degradation of
mammalian 3-hydroxy-3-methylglutaryl-coenzyme A reductase.
J Biol Chem 2000, 275:35840-35847.
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between the unfolded protein response and ER-associated 3:93-96.
degradation. Cell 2000, 101:249-258. See annotation Jiang et al. (2001) [41].
This paper and Friedlander et al. (2000) [23] make the very important
43. Murata S, Minami Y, Minami M, Chiba T, Tanaka K: CHIP is a
observation that the HRD ligase is required for normal cells to maintain an
chaperone-dependent E3 ligase that ubiquitylates unfolded
acceptable level of unfolded proteins in the ER, and that this machinery is
upregulated as one tactic to manage increases in the burden of lumenal protein. EMBO Rep 2001, 2:1133-1138.
unfolded proteins. See annotation Jiang et al. (2001) [41].

28. Braun S, Matuschewski K, Rape M, Thoms S, Jentsch S: Role of the 44. Meacham GC, Patterson C, Zhang W, Younger JM, Cyr DM:
ubiquitin-selective CDC48(UFD1/NPL4) chaperone (segregase) The Hsc70 co-chaperone CHIP targets immature CFTR for
in ERAD of OLE1 and other substrates. EMBO J 2002, proteasomal degradation. Nat Cell Biol 2001, 3:100-105.
21:615-621. This is a particularly relevant CHIP paper for this review. It demonstrates a
See annotation Bays et al. (2001) [11]. possible involvement of an adaptor ligase in the degradation of the misfolded
ER-membrane protein CFTR F508.
29. Rabinovich E, Kerem A, Frohlich KU, Diamant N, Bar-Nun S:
AAA-ATPase p97/Cdc48p, a cytosolic chaperone required for 45. Ponting CP: Proteins of the endoplasmic-reticulum-associated
endoplasmic reticulum-associated protein degradation. Mol Cell degradation pathway: domain detection and function prediction.
Biol 2002, 22:626-634. Biochem J 2000, 351:527-535.
See annotation Bays et al. (2001) [11].
46. Caldwell SR, Hill KJ, Cooper AA: Degradation of endoplasmic
30. Ye Y, Meyer HH, Rapoport TA: The AAA ATPase Cdc48/p97 and its reticulum (ER) quality control substrates requires transport
partners transport proteins from the ER into the cytosol. Nature between the ER and Golgi. J Biol Chem 2001, 276:23296-23303.
2001, 414:652-656. See annotation Vashist et al. (2001) [26].
See annotation Bays et al. (2001) [11].
47. Taxis C, Vogel F, Wolf D: ERGolgi-traffic is a prerequisite for
31. Rape M, Hoppe T, Gorr I, Kalocay M, Richly H, Jentsch S: efficient ER degradation. Mol Biol Cell 2002, in press.
Mobilization of processed, membrane-tethered SPT23 See annotation Vashist et al. (2001) [26].
transcription factor by CDC48(UFD1/NPL4), a ubiquitin-selective
chaperone. Cell 2001, 107:667-677. 48. Gardner RG, Hampton RY: A highly conserved signal controls
See annotation Bays et al. (2001) [11]. degradation of 3-hydroxy-3-methylglutaryl-coenzyme A
(HMG-CoA) reductase in eukaryotes. J Biol Chem 1999,
32. Jarosch E, Taxis C, Volkwein C, Bordallo J, Finley D, Wolf DH, 274:31671-31678.
Sommer T: Protein dislocation from the ER requires
polyubiquitination and the AAA-ATPase Cdc48. Nat Cell Biol 2002, 49. Cronin SR, Khoury A, Ferry DK, Hampton RY: Regulation of
4:134-139. HMG-CoA reductase degradation requires the P-type ATPase
See annotation Bays et al. (2001) [11]. Cod1p/Spf1p. J Cell Biol 2000, 148:915-924.

33. Hitchcock AL, Krebber H, Frietze S, Lin A, Latterich M, Silver PA: 50. Cronin S, Rao R, Hampton RY: Cod1p/Spf1 is an ER P-type
The conserved npl4 protein complex mediates proteasome- ATPase required for ER function and calcium homeostasis. J Cell
dependent membrane-bound transcription factor activation. Mol Biol 2002, in press.
Biol Cell 2001, 12:3226-3241.
See annotation Bays et al. (2001) [11]. 51. Gardner RG, Shan H, Matsuda SP, Hampton RY: An oxysterol-
derived positive signal for 3-hydroxy-3-methylglutaryl-CoA
34. Bays NW, Hampton RY: Cdc48p/Npl4p/Ufd1p: stuck in the middle reductase degradation in yeast. J Biol Chem 2001,
with Ub. Curr Biol 2002, 12:R366-R371. 276:8681-8694.
35. Plemper RK, Bohmler S, Bordallo J, Sommer T, Wolf DH: Mutant 52. Meigs TE, Simoni RD: Farnesol as a regulator of HMG-CoA
analysis links the translocon and BiP to retrograde protein reductase degradation: characterization and role of farnesyl
transport for ER degradation. Nature 1997, 388:891-895. pyrophosphatase. Arch Biochem Biophys 1997, 345:1-9.
482 Membranes and organelles

53. Correll CC, Ng L, Edwards PA: Identification of farnesol as the F508 cystic fibrosis transmembrane conductance regulator
non-sterol derivative of mevalonic acid required for the protein. Cell Stress Chaperones 1996, 1:117-125.
accelerated degradation of 3-hydroxy-3-methylglutaryl-coenzyme
A reductase. J Biol Chem 1994, 269:17390-17393. 57. Welch WJ, Brown CR: Influence of molecular and chemical
chaperones on protein folding. Cell Stress Chaperones 1996,
54. Gardner RG, Hampton RY: A distributed degron allows regulated entry 1:109-115.
into the ER degradation pathway. EMBO J 1999, 18:5994-6004.
58. Cheng HH, Xu L, Kumagai H, Simoni RD: Oligomerization state
55. Sato S, Ward CL, Krouse ME, Wine JJ, Kopito RR: Glycerol reverses influences the degradation rate of 3-hydroxy 3-methylglutaryl-
the misfolding phenotype of the most common Cystic fibrosis CoA reductase. J Biol Chem 1999, 274:17171-17178.
mutation. J Biol Chem 1996, 271:635-638.
59. Nohturfft A, Yabe D, Goldstein JL, Brown MS, Espenshade PJ:
56. Brown CR, Hong-Brown LQ, Biwersi J, Verkman AS, Welch WJ: Regulated step in cholesterol feedback localized to budding of
Chemical chaperones correct the mutant phenotype of the delta SCAP from ER membranes. Cell 2000, 102:315-323.