Evidence that this rule is WRONG

=================================================================
Direct Ubiquitination by  (rUbiq + pUbe1 + (rUbc13 + rUev1a))
=================================================================

  *** rTraf6[Ab] upshift[WBMS] is unchanged by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** source: 19675569-Fig-4a
  *** rTraf6[Ab] upshift[WBMS] is unchanged by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** source: 19675569-Fig-S2
  *** rTraf6[tAb] upshift[WBMS] is unchanged by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** source: 19675569-Fig-S2

  *** rTraf6[Ab]IP upshift[WBMS] is unchanged by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** comment: lysates were heated in 1% SDS to remove noncovalently attached proteins before IP
    *** source: 19675569-Fig-4b
  *** rTraf6[Ab]IP ubiq[UbAb] is unchanged by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** comment: lysates were heated in 1% SDS to remove noncovalently attached proteins before IP
    *** source: 19675569-Fig-4b

=================================================================
SemiDirect Ubiquitination by (rUbiq + pUbe1 + (rUbc13 + rUev1a))
===================================================================
Check out the Mobility Shifts of anti-Traf6 blots vs anti-Ub blots
notice that none of these were boiled before IP to remove non-covalently bound proteins
it is likely that the Ubiq staining seen is free polymerized Ubiq
see 19675569 and rule 080c
===================================================================


  *** xTraf6[tAb]IP ubiq[UbAb] is increased by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** IPfrom: HEK293 in BMS
    *** unaffected by: rUbiq(K63only) [substitution]
    *** source: 17135271-Fig-1b
  *** xTraf6[tAb]IP ubiq[tUbAb] is increased by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** inhibited by: xTraf6(C86A/H87A) [substitution]
    *** source: 19112497-Fig-S1
  *** xTraf6[tAb]IP upshift[WBMS] is increased by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** IPfrom: HEK293T in BMS
    *** inhibited by: xTraf6(C70A)[tAb]IP [substitution]
    *** inhibited by: xTraf6(mnr)"K32-348R"[tAb]IP [substitution]
    *** source: 19112497-Fig-2b
  *** xTraf6[tAb]IP upshift[WBMS] is increased by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** IPfrom: mEFs<Traf6~null> in BMS
    *** inhibited by: xTraf6(C70A)[tAb]IP [substitution]
    *** inhibited by: xTraf6(mnr)"K32-518R"[tAb]IP [substitution]
    *** source: 19112497-Fig-3b

===============================================================
Traf6 Ubiquitination by Ubc13/Uev1a produces K63 Ubiq linkages
===============================================================

  *** xTraf6[tAb]IP upshift[WBMS] is increased by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** IPfrom: HEK293T in BMS
    *** inhibited by: rUbiq(dig)"K48-only" [substitution]
    *** enhanced by: rUbiq(K63only) [substitution]
    *** source: 19112497-Fig-2b
  *** rTraf6[GST] ubiq[UbAb] is increased by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** inhibited by: xTraf6(C70A)"E3-mut" [substitution]
    *** inhibited by: rUbiq(K63R) [substitution]
    *** unaffected by: rUbiq(K63only) [substitution]
    *** unaffected by: rUbiq(K48R) [substitution]
    *** inhibited by: rUbiq(mnr)"K48-only" [substitution]
    *** source: 17135271-Fig-1c


  *** xTraf6((1-358)/GyrB)[tAb]IP ubiq[UbAb] is increased by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** inhibited by: rUbiq(dig)"no-lysines" [substitution]
    *** inhibited by: rUbiq(K63R) [substitution]
    *** enhanced by: rUbiq(K63only) [substitution]
    *** source: 11460167-Fig-4f
  *** xTraf6((1-358)/GyrB)[tAb]IP upshift[WBMS] is increased by (rUbiq + pUbe1 + rUbc13 + rUev1a)
    *** cells: none
    *** inhibited by: rUbiq(dig)"no-lysines" [substitution]
    *** inhibited by: rUbiq(K63R) [substitution]
    *** enhanced by: rUbiq(K63only) [substitution]
    *** source: 11460167-Fig-4f

================================
Expressed Traf6 is Ubiquitinated
================================

  *** xTraf6[Ab] upshift[WBMS] is detectable
    *** cells: HEK293 in BMS
    *** unaffected by: xTab2 [addition]
    *** source: 15327770-Fig-5a

reqs Ube2l3--

  *** xTraf6[tAb]IP ubiq[UbAb] is detectable
    *** cells: HEK293T in BMS
    *** comment: lysates were boiled to remove noncovalently attached proteins before IP
    *** reqs: Ube2l3 [RNAi]
    *** source: 19843958-Fig-S7b

does not req Ubc13--

  *** xTraf6[tAb]IP ubiq[UbAb] is detectable
    *** cells: HEK293T in BMS
    *** comment: lysates were boiled to remove noncovalently attached proteins before IP
    *** does not req: Ubc13 [RNAi]
    *** source: 19843958-Fig-S7b

Traf6 required features: C70 in Ring-type Zinc finger
                         K124R ubiquitination site

  *** xTraf6[tAb]IP ubiq[UbAb] is detectable
    *** cells: mEFs in BMLS
    *** inhibited by: xTraf6(C70A)"E3-mut" [substitution]
    *** partially inhibited by: xTraf6(K124R) [substitution]
    *** unaffected by: xTraf6(K133R) [substitution]
    *** source: 17135271-Fig-6a
  *** xTraf6[tAb]IP ubiq[tUbAb] is detectable
    *** cells: HEK293<xUbiq> in BMS
    *** inhibited by: xTraf6(C70A)"E3-mut" [substitution]
    *** source: 17135271-Fig-1a

