SCUD (Saccharomyces Cerevisiae Ubiquitination Database)


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Substrate Information

General Information
Standard nameMET4
Systematic nameYNL103W
DescriptionTranscriptional activator of sulfur metabolism MET4

Ubiquitination Features
NumberSiteTypeE2E3 ClassE3 SubclassSubstrate ReceptorUbiquitin ReceptorDUBEffect
1  CDC34RINGSCFMET30  Proteasomal Degradation
2  CDC34RINGSCFMET30  Proteolysis-independent Regulation
3  CDC34RINGSCFMET30   
4K163MULTI(K48-linked)CDC34RINGSCFMET30  Proteolysis-independent Regulation
5         
6         
* Blank: not determined, NA: not applicable.

References
[1]EvidenceProtein Stability Assay
MethodWestern blot
Reference"Feedback-regulated degradation of the transcriptional activator Met4 is triggered by the SCF(Met30 )complex."
Rouillon A, Barbey R, Patton EE, Tyers M, Thomas D.
EMBO J. 19(2):282-94 (2000) [10637232]
[2]EvidenceUb-conjugate Confirmation
MethodWestern blot
Reference"Regulation of transcription by ubiquitination without proteolysis: Cdc34/SCF(Met30)-mediated inactivation of the transcription factor Met4."
Kaiser P, Flick K, Wittenberg C, Reed SI.
Cell. 102(3):303-14 (2000) [10975521]
[3]EvidenceUb-conjugate Confirmation
MethodWestern blot
Reference"Dual regulation of the met4 transcription factor by ubiquitin-dependent degradation and inhibition of promoter recruitment."
Kuras L, Rouillon A, Lee T, Barbey R, Tyers M, Thomas D.
Mol Cell. 10(1):69-80 (2002) [12150908]
[4]Evidence 
Method 
Reference"Proteolysis-independent regulation of the transcription factor Met4 by a single Lys 48-linked ubiquitin chain."
Flick K, Ouni I, Wohlschlegel JA, Capati C, McDonald WH, Yates JR, Kaiser P.
Nat Cell Biol. 6(7):634-41 (2004) [15208638]
[5]EvidencePurification & Protein ID
Methodpurification (HB-ubiquitin (a tandem affinity tag under denaturing conditions)), LC/MS
Reference"A tandem affinity tag for two-step purification under fully denaturing conditions: application in ubiquitin profiling and protein complex identification combined with in vivocross-linking."
Tagwerker C, Flick K, Cui M, Guerrero C, Dou Y, Auer B, Baldi P, Huang L, Kaiser P.
Mol Cell Proteomics. 5(4):737-48 (2006) [16432255]
[6]EvidencePurification & Protein ID
Methodpurification (6xHis-Myc-ubiquitin), LC/MS, validation (virtual Western blot)
Reference"Systematic approach for validating the ubiquitinated proteome."
Seyfried NT, Xu P, Duong DM, Cheng D, Hanfelt J, Peng J.
Anal Chem. 80(11):4161-9 (2008) [18433149]

Cross References
Swiss-Prot accessionP32389
SGD YNL103W
Gene Ontology (GO)
Cellular componentGO:0005634nucleusIPI:SGD.
Molecular functionGO:0003713transcription coactivator activityIDA:SGD.
Biological processGO:0045944positive regulation of transcription from R...IDA:SGD.
Biological processGO:0046685response to arsenicIDA:SGD.
Biological processGO:0046686response to cadmium ionIDA:SGD.
Biological processGO:0000096sulfur amino acid metabolic processIMP:SGD.
EC number  

Additional Features
Other Post Translational Modifications (PTM)
Residue indexType
206Phosphothreonine.
302Phosphothreonine.
307Phosphoserine.
310Phosphoserine.
564Phosphoserine.

Sequence Information
Sequence length672
Molecular weight74373

	---------+----------+----------+----------+----------+
MKQEQSHEGD SYSTEFINLF GKDTATHPSS NNGANNNGMG STNSLDQFVA   50
TASSSSSLVT SSENRRPLIG DVTNRGNTNL YDHAVTPEIL LEQLAYVDNF   100
IPSLDNEFSN VDWNVNTTHN NANNNGADTF SSINANPFDL DEQLAIELSA   150
FADDSFIFPD EDKPSNNNNN SNNGNDDHSN HDVLHEDPST NNRQRNPHFL   200
TQRRNTFLTS QYDQSKSRFS SKNKRNGNNG ETNNFGDNMQ NNHPFEPNFM   250
GSPSQFPADA TNMTSIDHGG FTNVDITSTE NNTTGDNGVD ALSNLLHRTT   300
HTPNRSSPLS NVTSAQNSSS QQRKHSESKV DSNSDNNSSN KAPNITVPDY   350
SIIPTSVLVT LLPRVNVPNG AYNSLISAGF DNDQIDAIAA IMAYHHQKKI   400
RENNSNNNKN INTNDSQEAP ILKNINELLS VLIPPSPAET RGPTTLSTSP   450
SFNEHGVVAE ASFLSSILEL GIKHPKSNNI HNQRQPSRND HKISRESDGN   500
NGNDNVHHNN AVIKSSTTRG DEIAKIRSEP TLNASSSDHK ENSLKRSHSG   550
DLKNKKVPVD RKYSDNEDDE YDDADLHGFE KKQLIKKELG DDDEDLLIQS   600
KKSHQKKKLK EKELESSIHE LTEIAASLQK RIHTLETENK LLKNLVLSSG   650
ETEGIKKAES LKKQIFEKVQ KE                                 672

	* Ubiquitinated lysine is denoted as red color.