A telomerase with novel non-canonical roles: TERT controls cellular aggregation and tissue size inDictyostelium

AbstractTelomerase, particularly its main subunit, the reverse transcriptase, TERT, prevents DNA erosion during eukaryotic chromosomal replication, but also has poorly understood non-canonical functions. Here, in the model social amoebaDictyostelium discoideum,we show that the protein encoded byterthas telomerase-like motifs and regulates, non-canonically, important developmental processes. Expression levels of wild-type (WT)tertwere biphasic, peaking at 8 and 12 h post-starvation, aligning with developmental events, such as the initiation of streaming (∼7 h) and mound formation (∼10 h). IntertKO mutants, however, aggregation was delayed until 16 h. Large, irregular streams formed, then broke up, leading to small mounds. The mound-size defect was not induced when a KO mutant ofcountin(a master size-regulating gene) was treated with TERT inhibitors but anti-countin antibodies did rescue size in thetertKO. Further, conditioned medium fromcountinmutants failed to rescue size in thetertKO, but the converse experiment worked. These and additional observations indicate that TERT acts upstream ofsmlA/countinto regulate tissue size: (i) the observed expression levels ofsmlAandcountin,being respectively lower and higher (than WT) in thetertKO; (ii) the levels of known size-regulation intermediates, glucose (low) and adenosine (high), in thetertmutant, and the size defect’s rescue by supplementing glucose or the adenosine-inhibitor, caffeine; (iii) the induction of the size defect in the WT bytertKO conditioned medium and TERT inhibitors. ThetertKO’s other defects (delayed aggregation, irregular streaming) were associated with changes to cAMP-regulated processes (e.g. chemotaxis, cAMP pulsing) and their regulatory factors (e.g. cAMP;acaA, carAexpression). Overexpression of WTtertin thetertKO rescued these defects (and size), and restored a single cAMP signalling centre. Our results indicate that TERT acts in novel, non-canonical and upstream ways, regulating key developmental events inDictyostelium.Author summaryWhen cells divide, their chromosomes are prone to shrinkage. This risk is reduced by an enzyme that repairs protective caps on each chromosome after cell division. This enzyme, telomerase, also has several other important but unrelated roles in human health. Most importantly, via one or other of its functions, both high and low levels of the enzyme can contribute to cancer. We have studied, for the first time, the roles played by telomerase in the life-cycle of the cellular slime mould,Dictyostelium discoideum, a model system with a rich history of helping us understand human biology. While we did not find any evidence of telomerase having the features typically needed to repair a chromosome, telomerase was necessary for many aspects of development. In forming the fruiting bodies that helpDictyosteliumreproduce, a mutant that lacks telomerase miscalculates how big those bodies should be, and they end up being too small. Also, earlier, during an earlier stage, aggregation, the migration of cells that form each fruiting body is delayed and irregular. These results are significant because they show, for the first time, that a telomerase can influence cell migration and tissue size regulation, two processes involved in a wide range of cancers.