Ubiquitin-mediated proteolysis

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Contents

Related enzymes in Selaginella moellendorffii

Ying Li (li23@purdue.edu) and Michael Gribskov (gribskov@purdue.edu)

Ubiquitin-activating enzyme (UBAs, or E1s), Ubiquitin-conjugating enzymes (UBCs, or E2s) and Ubiquitin-protein ligases (E3s) are the three types of enzymes involved in the ubiquitin-mediated proteolysis process in plants/animals. Proteins destined to be degraded are multi-ubiquitinated through a process catalyzed by the above enzymes, then are recognized and degraded by the 26S proteasome.

There are fewer E1 isoforms compared to E2 isoforms and the much more complicated E3 isoforms in plants (see Table 1, Table 2 and Table 4). Eighteen E2s are identified based on protein sequences comparison with other plant species. Table 2 shows the grouping based on sequence similarity (Kraft, et al., 2005) and comparison of UBCs among Arabidopsis thaliana, Oryza sativa , Selaginella moellendorffii and Physcomitrella Patens. Table 3 lists the number of putative E3s (ordered by E3 subfamilies) in Selaginella moellendorffii.

Table 1. Comparison of E1s

E1 Number of putative orthologs*
Arabidopsis Oryza Selaginella Physcometrella
UBA1/UBA2 2 4 1 (2) 2

Table 2. Grouping and comparison of E2s

Group E2 Number of putative orthologs*
Arabidopsis Oryza Selaginella Physcomitrella
I SCE1a 1 1 1 1
II RCE1 1 0 0 0
II RCE2 1 1 (3) 2 (2,2) 2 (2,3)
III UBC1 1 1 (3) 2 (2,2) 1 (2)
III UBC2 1 0 0 0
III UBC3 1 0 0 0
IV UBC4 1 1 (1) 0 1 (1)
IV UBC5 1 0 2 (2,2) 1 (1)
IV UBC6 1 0 0 0
V UBC7 1 1 (1) 0 0
V UBC13 1 1 (1) 1 (2) 1 (3)
V UBC14 1 0 0 1 (1)
VI UBC8 1 1 (1) 1 (2) 0
VI UBC9 1 1 (1) 0 1 (1)
VI UBC10 1 1 (3) 0 0
VI UBC11 1 1 (1) 1 (2) 0
VI UBC12 1 0 0 0
VI UBC28 1 1 (1) 1 (4) 1 (3)
VI UBC29 1 0 0 0
VI UBC30 1 0 0 1 (2)
VII UBC15 1 1 (1) 1 (2) 1 (2)
VII UBC16 1 0 1 (2) 1 (2)
VII UBC17 1 0 0 0
VII UBC18 1 1 (1) 0 0
VIII UBC19 1 0 1 (2) 0
VIII UBC20 1 1 (1) 0 1 (1)
IX UBC21 1 1 (1) 0 1 (1)
X UBC22 1 1 (1) 1 (2) 1 (1)
XI UBC23 1 1 (2) 0 1 (1)
XI UBC24 1 1 (1) 0 0
XI UBC25 1 1 (5) 0 0
XI UBC26 1 1 (1) 0 0
XII UBC27 1 1 (1) 1 (2) 1 (1)
XIII UBC31 1 0 0 0
XIV UBC32 1 1 (1) 1 (2) 1 (1)
XIV UBC33 1 1 (1) 1 (2) 1 (1)
XIV UBC34 1 0 0 0
XV UBC35 1 1 (1) 1 (2) 1 (1)
XV UBC36 1 0 0 1 (1)
XVI UBC37 1 1 (1) 1 (2) 1 (1)
Total 37 23 17 21

footnote: number of putative loci( number of putative alleles detected, separated by comma if there are more than one group of alleles).

Table 3. Gene models in the assembly

protein id gene name named by model changes
271478 UBC1-1 YING LI no
228869 UBC1-2 YING LI no
228475 UBC1-3 YING LI no
228312 UBC1-4 YING LI no
450658 UBC5-1 YING LI yes
450660 UBC5-2 YING LI yes
81550 UBC5-3 YING LI no
174924 UBC5-4 YING LI no
74865 UBC13-1 YING LI no
87352 UBC13-2 YING LI no
449723 UBC8-1 GRIBSKOV yes
450670 UBC8-2 YING LI yes
187468 UBC11-1 YING LI yes
233935 UBC11-2 YING LI yes
449724 UBC28-1 GRIBSKOV yes
228916 UBC28-2 YING LI no
270923 UBC28-3 YING LI no
270384 UBC28-4 YING LI no
270987 UBC15-1 YING LI no
159187 UBC15-2 YING LI no
268066 UBC16-1 YING LI no
172555 UBC16-2 YING LI no
181155 UBC19-1 YING LI no
450635 UBC19-2 YING LI yes
145850 UBC22-1 YING LI no
160563 UBC22-2 YING LI no
186167 UBC27-1 YING LI no
95808 UBC27-2 YING LI no
450643 UBC32-1 YING LI yes
450644 UBC32-2 YING LI yes
36529 UBC33-1 YING LI no
15877 UBC33-2 YING LI no
116003 UBC35-1 YING LI no
134879 UBC35-2 YING LI no
5749 UBC37-like-1 YING LI no
6339 UBC37-like-2 YING LI no


Table 4. Number of putative E3 Orthologs in Arabidopsis thalinana and Selaginella moellendorffii

E3 subfamily Number in ATH1 Best Arabidopsis protein sequences match in Selmo1 Best Selaginella protein sequences mathc in ATH1 Arabidopsis-Selaginella sequence match combination
ASK 21 4 4 8
BTB 80 33 40 66
Cullin 10 5 8 10
HECT 7 6 8 12
UBOX 60 19 25 36
FBOX 679 98 154 192
RingFinger 470 161 204 306
PHD 215 52 68 101
DDB 5 4 8 8
RBX1 2 0 0 0

Note1: The matches include 1(arabidopsis gene)-to-2(selaginella alleles) and 2(arabidopsis genes)-to-1(selaginella allele) relationship, e.g., for ASK, each of the unique four arabidopsis protein sequences matched to two selaginella allele sequences, representing a 1-to-1 gene relationship; each of the selaginella allele matches to two candidate arabidopsis genes (which should be finalized to one eventually). The above relationship results in a combination of 8 matches.

Note2: Steps for obtaining the above results.

  • Step 1: Run blastp using different queries and database (Blastp results are listed in tabular format)
    • use Selmo1 protein sequences as the queries and ATH1 E3 protein sequences as the database
    • use ATH1 E3 protein sequences as the queries and Selmo1 protein sequences as the database
  • Step 2: Get common match sequences
    • obtain common match sequences joined on Arabidopsis gene name (or AGI) and Selaginella protein ID.
  • Step 3: Sort by E-values (smallest to biggest)
    • Run a perl script that takes the common match sequences from step 2 and sorts E-values by Arabidopsis gene name and Selaginella protein ID, respectively. Output two corresponding lists.
  • Step 4: Get the top two matches by selecting the smallest two E-values
    • Join the two list by Arabidopsis gene name and Selaginella protein ID and filter the results using criteria (E-value rank = 1 or 2 for both ranks in the two lists).

Reference

Kraft,E., S.L. Stone, L. Ma, N. Su, Y. Gao, O. Lau, X. Deng, and J. Callis, Genome Analysis and Functional Characterization of the E2 and RING-Type E3 Ligase Ubiquitination Enzymes of Arabidopsis. Plant Physiol. 2005 December; 139(4): 1597–1611.

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