For evaluation purposes, we used a trimmed version of this reference where the terminal copies of RL and RS have been removed, leaving a sequence of 136 kb.
Curiously, the C-terminal copy of the catalytic domain has lost the catalytically important HIGH and KMSKS motifs characteristic of class I aminoacyl-tRNA synthetases.
Besides an extra C-terminal copy of the ancestral hydroperoxidase gene resulting from gene duplication, these two large loops are likely to control the orientation of both the haem group and of essential residues in the active site.
Interestingly, it can be seen that it is the N-terminal copy of SUMO containing the His6 tag, that accepts the ubiquitin in these reactions.
Interestingly, this change affects the two youngest, 3′-terminal copies in the tandem gene clusters (Song et al. 2001).
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Two smaller grn genes, grn1 and grn2, each encoding one full and one amino-terminal half copies only of the granulin motif, were also characterized.
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As such, a mutagenesis study was undertaken using a new peptide-linked poly-SUMO-2 within which only the N-terminal SUMO-2 copy contained an intact N-terminus, with the remaining three copies comprising residues 12 92.
Our own examination of the heterothallic O. ulmi W9 genome shows that it has an N-terminal-truncated nonfunctional copy of MAT1-1-1 in its MAT1-2 idiomorph, as do many other mating-type-2 Sordariomycetes (fig. 7).
Log phase T. brucei procyclic strain 449 cells were transfected with BsmI-digested pTbARL6PTP by nucleofection as in Section 2.7, replacing one endogenous allele of ARL6 with a C-terminal PTP tagged copy of the ARL6 open reading frame.
