Table I. In vitro phosphorylation of potential cyclin E–Cdk2 substrates by different cyclin–Cdk complexes.
| Potential cyclin E–Cdk2 substrate
|
Phosphorylation sites and cyclin binding motifs in the RZPD clone |
Phosphorylation of the RZPD clone by kinasesa |
||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Gene | ORF of RZPD clone |
Name/biological process | min: S/TP |
cons: S/TPXK/R |
RXL | D1–K4 | D3–K4 | E–K2 | A–K2 | B–K1 |
| NUMA1b | 245 aa C-term |
Nuclear mitotic apparatus protein 1/ spindle apparatus organization |
7 | 1 | 0 | − | ± | + | + | − |
| LOC201191 | 231 aa C-term |
Hypothetical protein LOC201191/unknown |
6 | 1 | 2 | − | ± | + | + | − |
| PIP5K1C | 200 aa C-term |
Phosphatidylinositol-4-phosphate 5 kinase, type I γ/phosphatidylinositol signaling |
7 | 2 | 1 | ± | + | + | + | ± |
| CCNL2 | 136 aa C-term |
cyclin L2 / RNA processing | 7 | 3 | 2 | − | − | ± | + | ± |
| RASL11B | 118 aa C-term |
RAS-like, family 11, member B/signaling? | 2 | 1 | 0 | − | + | + | + | − |
| SIRT2 | full-lengthc | Sirtuin (silent mating type information regulation 2 homologue) 2 (S. cerevisiae)/ NAD+-dependent histone/protein deacetylase |
3 | 1 | 1 | − | − | + | + | ± |
| FLJ13111 | full-length | Hypothetical protein FLJ13111/unknown | 4 | 2 | 4 | − | ± | + | + | ± |
| SUPT5Hb | 120 aa C-term |
Suppressor of Ty 5 homologue (S. cerevisiae)/transcription |
1 | 1 | 1 | − | − | ± | − | − |
| STMN2b | full-length | Stathmin-like 2/microtubule destabilization in neuronal growth |
2 | 1 | 1 | − | + | + | + | − |
| LOC146909 | 114 aa C-term |
Hypothetical protein LOC146909/unknown |
1 | 0 | 1 | − | + | + | + | − |
| EEF1G | 211 aa C-term |
Eukaryotic translation elongation factor 1γ/translation |
2 | 1 | 1 | − | ± | + | + | ± |
| TRIT1 | 99 aa C-term |
tRNA isopentenyltransferase 1/ RNA modification |
3 | 1 | 1 | − | + | + | + | ± |
| LOC388799 | 89 aa C-term |
Hypothetical protein LOC388799/unknown |
2 | 1 | 0 | − | ± | + | + | ± |
| STUB1 | full-length | STIP1 homology and U-box–containing protein 1/protein turnover |
4 | 1 | 3 | − | + | + | + | ± |
| SRRM2 | 98 aa central |
Serine/arginine repetitive matrix 2/ RNA processing |
3 | 0 | 0 | − | − | + | + | + |
| TALDO1 | full-length | Transaldolase 1/metabolism | 3 | 1 | 6 | − | + | + | + | − |
| SFRS1 | 243 aa C-term |
Splicing factor, arginine/serine-rich 1 (splicing factor 2, alternate splicing factor)/RNA processing |
4 | 1 | 1 | − | + | + | + | − |
| SNAPAP | full-length | SNAP-associated protein/exocytosis of synaptic vesicles |
2 | 1 | 2 | + | ± | + | ||
| LOC339287 | 90 aa central |
Hypothetical protein LOC339287/unknown |
5 | 4 | 1 | + | + | + | + | + |
| FLJ12949 | 248 aa C-term |
Hypothetical protein FLJ12949/unknown | 4 | 1 | 3 | − | + | + | − | − |
| MAP2 | 376 aa C-term |
MAP2/microtubule assembly in neurogenesis |
16 | 3 | 1 | − | + | + | + | − |
| RPL14 | 211 aa C-term |
Ribosomal protein L14/translation | 1 | 1 | 1 | − | ± | − | − | − |
| FNBP3 | 245 aa C-term |
Formin binding protein 3/splicing? | 2 | 1 | 1 | − | ± | ± | ± | − |
| PRC1d | full-length | Protein regulating cytokinesis 1/ cytokinesis |
5 | 2 | 2 | − | + | + | + | ± |
| FLJ13305 | 162 aa central |
Hypothetical protein FLJ13305/unknown | 4 | 0 | 3 | − | + | + | ± | − |
| ING5 | full-lengthc | Inhibitor of growth family, member 5/ p53 pathway, replication |
2 | 1 | 3 | − | − | + | + | − |
a
The kinases used are abbreviated (e.g., D1–K4 for cyclin D1–Cdk4). The substrates are phosphorylated strongly (+), weakly (±), or not at all (−) by the respective kinase.
b
Previously reported Cdk substrate (Stachora et al., 1997; Gavet et al., 1998; Sun and Schatten, 2006).
c
For ING5 and SIRT2, GST-tagged full-length proteins were used in the kinase assays.
d
Previously reported cyclin E–Cdk2 substrate (Jiang et al., 1998).