Fine-Mapping, Genomic Organization, and Transcript Analysis of the Human Ubiquitin-Conjugating Enzyme Gene UBE2L3

Moynihan, Terry P.; Cole, Charlotte G.; Dunham, Ian; O'Neil, Lisa; Markham, Alexander F.; Robinson, Philip A.

doi:10.1006/geno.1998.5257

Cited by 21 publications

(16 citation statements)

References 14 publications

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“…The "bait" plasmid was constructed by cloning the open reading frame of UbcH7 (13,20,21) (CLONTECH). Clones were selected on minimal synthetic dropout medium in the absence of tryptophan.…”

Section: Methodsmentioning

confidence: 99%

“…Interaction of HHARI with Human E2s in Vitro-Four human E2s were assessed for their capacity to form a stable association with HHARI: (i) UbcH1 (24), which does not interact with E6-AP, (ii) UbcH5 (25), (iii) UbcH7 (13,20,21), and (iv) UbcH8 (14), all of which support E6-AP-ubiquitin thioester formation. Plasmids bearing upstream bacteriophage T7 promoters encoding these E2s have been described previously (6,13,25).…”

Section: Methodsmentioning

confidence: 99%

“…Identification of UbcH7 Domains Regulating the Interaction with HHARI-Specific regions of the UbcH7 cDNA (15,20,21) were amplified by PCR using Pfu DNA polymerase and cloned into the BamHI site of pET32a as in-frame thioredoxin (TXD) C-terminal fusion proteins (Novagen). Binding assays were performed as described above using ϳ0.5 g each of TXD fusion protein and GST-HHARI (118 -557).…”

Section: Interaction Of Gst-hhari (118 -557) With Endogenous Ubch7 Inmentioning

confidence: 99%

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The Ubiquitin-conjugating Enzymes UbcH7 and UbcH8 Interact with RING Finger/IBR Motif-containing Domains of HHARI and H7-AP1

Moynihan

Ardley

Nuber

et al. 1999

Journal of Biological Chemistry

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110

104

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Ubiquitinylation of proteins appears to be mediated by the specific interplay between ubiquitin-conjugating enzymes (E2s) and ubiquitin-protein ligases (E3s). However, cognate E3s and/or substrate proteins have been identified for only a few E2s. To identify proteins that can interact with the human E2 UbcH7, a yeast twohybrid screen was performed. Two proteins were identified and termed human homologue of Drosophila ariadne (HHARI) and UbcH7-associated protein (H7-AP1). Both proteins, which are widely expressed, are characterized by the presence of RING finger and in between RING fingers (IBR) domains. No other overt structural similarity was observed between the two proteins. In vitro binding studies revealed that an N-terminal RING finger motif (HHARI) and the IBR domain (HHARI and H7-AP1) are involved in the interaction of these proteins with UbcH7. Furthermore, binding of these two proteins to UbcH7 is specific insofar that both HHARI and H7-AP1 can bind to the closely related E2, UbcH8, but not to the unrelated E2s UbcH5 and UbcH1. Although it is not clear at present whether HHARI and H7-AP1 serve, for instance, as substrates for UbcH7 or represent proteins with E3 activity, our data suggests that a subset of RING finger/IBR proteins are functionally linked to the ubiquitin/proteasome pathway.The primary role of the protein ubiquitinylation pathway is the targeting of intracellular substrate proteins for degradation (1). In this process, ubiquitin is first activated in an ATPdependent step forming a thioester bond with an ubiquitinactivating enzyme (E1). Ubiquitin is then transferred to an ubiquitin-conjugating enzyme (E2), 1 retaining the high energy thioester bond. Thereafter, the E2, alone or in conjunction with an ubiquitin-protein ligase (E3), catalyzes the final attachment of ubiquitin to the target protein (2). Ubiquitin itself can then serve as a ubiquitinylation substrate, resulting in the generation of polyubiquitinylated proteins possibly with the aid of an E4 (3). Finally, ubiquitinylated proteins are recognized and degraded by the 26 S proteasome. It has been proposed that the selection of an individual protein for proteasomal degradation via the ubiquitin pathway requires a unique combination of an E2 and E3. In S. cerevisiae, some 13 E2s or E2-related proteins have been identified, and many more are present in higher eukaryotes (2). E2s are characterized by a conserved catalytic domain of approximately 150 amino acid residues. Despite their functional redundancy, individual E2s appear to be involved in different cellular processes and, therefore, in the ubiquitinylation of different substrate proteins. The distinct substrate specificity of E2s is at least in part explained by the observation that different E2s interact with different E3s.Four classes of E3 have been identified to date; in contrast to E2s, they exhibit no overt sequence homology. These are yeast UBR1 and its mammalian homologues (4), mammalian E6-AP

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“…The "bait" plasmid was constructed by cloning the open reading frame of UbcH7 (13,20,21) (CLONTECH). Clones were selected on minimal synthetic dropout medium in the absence of tryptophan.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Interaction Of Gst-hhari (118 -557) With Endogenous Ubch7 Inmentioning

confidence: 99%

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The Ubiquitin-conjugating Enzymes UbcH7 and UbcH8 Interact with RING Finger/IBR Motif-containing Domains of HHARI and H7-AP1

Moynihan

Ardley

Nuber

et al. 1999

Journal of Biological Chemistry

Self Cite

110

104

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“…This protein is involved in the ubiquitination and degradation of the oncogenes MYC 56 and c-FOS. 57 Therefore, UBE2L3 down-regulation in MF cases may assist MF tumorigenesis ( Figure 6). …”

Section: Tnfsf5 (Cd40l or Tnf-related Activation Protein [Trap]mentioning

confidence: 99%

Mycosis fungoides shows concurrent deregulation of multiple genes involved in the TNF signaling pathway: an expression profile study

Tracey

Villuendas

Dotor

et al. 2003

Blood

155

116

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Mycosis fungoides (MF) is the most frequent type of cutaneous T-cell lymphoma, whose diagnosis and study is hampered by its morphologic similarity to inflammatory dermatoses (ID) and the low proportion of tumoral cells, which often account for only 5% to 10% of the total tissue cells. cDNA microarray studies using the CNIO OncoChip of 29 MF and 11 ID cases revealed a signature of 27 genes implicated in the tumorigenesis of MF, including tumor necrosis factor receptor (TNFR)-dependent apoptosis regulators, STAT4, CD40L, and other oncogenes and apoptosis inhibitors. Subsequently a 6-gene prediction model was constructed that is capable of distinguishing MF and ID cases with unprecedented accuracy. This model correctly predicted the class of 97% of cases in a blind test validation using 24 MF patients with low clinical stages. Unsupervised hierarchic clustering has revealed 2 major subclasses of MF, one of which tends to include more aggressive-type MF cases including tumoral MF forms. Furthermore, signatures associated with abnormal immunophenotype (11 genes) and tumor stage disease IntroductionMycosis fungoides (MF) is a peripheral cutaneous T-cell lymphoma (CTCL) characterized by the infiltration and accumulation of tumoral T cells in the skin (epidermal and dermal infiltration). 1 It is the most frequent type of CTCL, representing more than half of all lymphomas originating in the skin. 2 Clinical and pathologic diagnosis of MF has proved very difficult since it bears many resemblances to common inflammatory dermatoses (ID) such as interface and spongiotic dermatitis, conditions involving infiltration and accumulation of benign T cells in the skin. The study of MF is further compounded by the small number of tumoral cells, which may be present in the skin, with tumoral T-cell infiltrate often accounting for only 5% to 10% of the total tissue cells. Most CTCLs have the phenotype of T-helper memory lymphocytes (CD3 ϩ , CD4 ϩ ) with only a minority of cases showing phenotypes such as CD4 Ϫ or CD8 ϩ , with CD8 ϩ cases possibly representing a subset of aggressive cases. 3 Additionally, loss of CD7 expression is considered a distinguishing characteristic of MF. 4 The etiology of MF remains largely unknown, although some triggering factors such as chronic antigen stimulation 5 or human T-cell lymphotropic virus type 1 or other viral infections have been proposed. However, some of these findings have been somewhat contentious. 6,7 Molecular studies of MF have revealed some interesting facts, although the studies are confined to small groups of genes and proteins. Defective FAS signaling has been suggested as a possible causative agent in MF pathogenesis due to defects in apoptosis signaling in skin-homing T cells. 8 Reduction of FAS expression has been documented in peripheral blood CD4 ϩ T lymphocytes in CTCL. 9 This decreased FAS expression may be the result of FAS mutations, which are infrequent but are present in some cases. 8 Defective FAS signaling may also be supplemented by mutations or defects in caspases or ...

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“…We have previously characterized the genomic structure of UBE2L3, the UbcH7 gene encoded by four exons (Moynihan et al, 1996(Moynihan et al, , 1998. Due to their apparent functional redundancy, and similarities in primary amino acid structure, we sought to establish whether the genomic structure of UBE2L6 was similar at the DNA level to that of UBE2L3 (Moynihan et al, 1996(Moynihan et al, , 1998.…”

Section: Ubch8mentioning

confidence: 99%

Genomic organization of the human ubiquitin-conjugating enzyme gene, UBE2L6 on chromosome 11q12

Ardley¹,

Rose²,

Tan³

et al. 2000

Cytogenet Genome Res

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The human UBE2L6 gene encodes UbcH8^Kumar, a ubiquitin-conjugating enzyme (E2) highly simliar in primary structure to UbcH7 which is encoded by UBE2L3. Like UBC4 and UBC5 in yeast, these proteins demonstrate functional redundancy. Herein we report the intron/exon structure of UBE2L6. Comparison of the genomic organization of UBE2L6 with UBE2L3 demonstrates that these genes remain highly conserved at the genomic as well as at the protein level. We also describe the chromosomal localization of UBE2L6, which maps to chromosome 11q12.

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Fine-Mapping, Genomic Organization, and Transcript Analysis of the Human Ubiquitin-Conjugating Enzyme Gene UBE2L3

Cited by 21 publications

References 14 publications

The Ubiquitin-conjugating Enzymes UbcH7 and UbcH8 Interact with RING Finger/IBR Motif-containing Domains of HHARI and H7-AP1

The Ubiquitin-conjugating Enzymes UbcH7 and UbcH8 Interact with RING Finger/IBR Motif-containing Domains of HHARI and H7-AP1

Mycosis fungoides shows concurrent deregulation of multiple genes involved in the TNF signaling pathway: an expression profile study

Genomic organization of the human ubiquitin-conjugating enzyme gene, UBE2L6 on chromosome 11q12

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