The following information accompanied stocks donated to the Bloomington Stock Center by Dirk Beuchle and Beat Suter, University of Bern.
TTLL5R339A: Point mutation = Arg (AGG) -> Ala (GCT) on the reverse strand; introducing an AluI site (AGCT) at the same time. Drosophila TTLL5R339 corresponds to mouse TTLL5R188, which is vital for TTLL5’s glutamylation activity. Mouse TTLL5R188 resides in the beta6-7 loop, which forms a salt-bridge interaction with the C-terminal tail of alpha-tubulin and orients it towards the ATP/ADP-binding site (van Dijk et al., 2007; Natarajan et al., 2017).
TTLL5E517A: Point mutation = Glu (GAA) -> Ala (GCC) on the reverse strand; introducing a HaeIII site (GGCC) at the same time. Drosophila TTLL5 residues E517 correspond to mouse E336, which directly interacts with ADP/ATP and thus affect general TTLL5 enzymatic activity in mouse (van Dijk et al., 2007; Natarajan et al., 2017).
TTLL5P522A: Point mutation = Pro (CCG) -> Ala (GCG) on the reverse strand; introducing a BsaHI site (GRCGYC) at the same time. Drosophila TTLL5P522 codon corresponding to human TTLP336 (and human TTLL5P371/mouse TTLL5P371), which is required for anchoring TTL to alpha-tubulin by forming hydrogen bonds with C-terminal tail residues of alpha-tubulin (Prota et al., 2013).
TTLL5MiEx: Generated by imprecise excision of Mi{MIC}TTLL5MI01917, which caused the formation of a premature stop codon at gene position 8,132 in the open reading frame (codon position 392).
UASp- venus::TTLL5:  The DNA was inserted into the pUASP-venus vector via Xba l/Bgl II ligation and targeted into the attP-58A and attP-64A landing platforms.
UASp- venus::TTLL5-E517A:  TTLL5 point mutation: E517A. Drosophila TTLL5 residues E517 correspond to mouse E336, which directly interacts with ADP/ATP and thus affect general TTLL5 enzymatic activity in mouse (van Dijk et al., 2007; Natarajan et al., 2017). Targeted into the attP-58A and attP-64A landing platforms.
UASp- venus::TTLL5-E517G:  TTLL5 point mutation: E517G. Drosophila TTLL5 residues E517 correspond to mouse E336, which directly interacts with ADP/ATP and thus affect general TTLL5 enzymatic activity in mouse (van Dijk et al., 2007; Natarajan et al., 2017). Targeted into the attP-58A and attP-64A landing platforms.