, 3769C3780. impaired the function of multiciliated epidermal cells in propelling planarian motion, as well as the osmoregulatory function of protonephridia. Decreased density and structural defects of motile cilia were observed in the epidermis of by phase contrast, immunofluorescence, and transmission electron microscopy. Altogether, these results demonstrate that members of the TTBK family of proteins are postmeiotic regulators of sperm development and also contribute to the formation of motile cilia in the soma. INTRODUCTION Ciliopathies are a group of health disorders associated with mutations of factors involved in cilia formation and function. Included among these are variable pathologies of the kidney, liver, and retina, as well as brain dysgenesis, neurocognitive impairments, and infertility (Badano is almost exclusively expressed in the CNS, and certain alleles are associated with late onset Alzheimers disease (Vazquez-Higuera expression is usually detected broadly at the transcript level, but the protein is particularly abundant in the brain and testis (Bouskila are associated with spinocerebral ataxia type 11, which is usually characterized by progressive loss of coordination, difficulty walking, abnormal eye signs, peripheral neuropathy, and dystonia (Houlden paralogues is usually highest in male gonads (Sato transgenic mice display phosphorylated neurofilament aggregation and Splitomicin age-dependent memory impairment (Sato mutant mice manifest defects in primary cilia formation, loss of Sonic Splitomicin hedgehog signaling activity, and ultimately die during embryogenesis Mouse monoclonal antibody to LIN28 (Bouskila function in mice, uncovering the function of TTBK phosphorylation during spermatogenesis may require analyses in nonmammalian organisms. Several studies have recently demonstrated the value of the planarian flatworm for in vivo evaluation of genes involved in cilia formation and function (Rompolas develop their entire hermaphroditic reproductive system postembryonically, offering the opportunity to analyze genetic contributions to the formation of gametes and ciliated structures of the reproductive system after embryonic development is usually completed (Zayas and preferentially expressed in testes A TBLASTN search revealed six sequences with partial identity to human TTBK1 and TTBK2 (E value 2.37e-73) among contigs from a reference transcriptome assembly of the sexual strain of (Rouhana genome sequences (Robb (Supplemental Table S1). Orthology to TTBK family proteins was confirmed using reciprocal BLASTP searches between predicted planarian translation products and human RefSeq protein sequences deposited in the National Center for Biotechnology Information (NCBI) (Supplemental Table S2). Planarian TTBKs share 61C71% protein sequence identity with human orthologues within the conserved kinase domain name, whereas 85% identity is usually shared between human paralogues in this region (Physique 1A). In addition, conserved TTBK1 active site residues (Kiefer (Supplemental Physique 1). No conservation was detected outside of the kinase domain name between planarian and human TTBK sequences. However, regions of less than 50% identity are shared between human paralogues outside of the kinase domain name, and a stretch smaller than 50 amino acids with 46% identity is usually shared between Smed-TTBK-c and Smed-TTBK-d (Physique 1A). These analyses suggest that all six planarian TTBK homologues are active kinases that derived from an ancestral TTBK sequence, and that human paralogues are a result of Splitomicin a Splitomicin recent duplication not ancestrally present before divergence of lophotrochozoans and deuterostomes. Furthermore, the lack of sequence conservation outside of the kinase domain name corroborates with observations made by Ikezu and Ikezu (2014) , who proposed that current TTBK homologues arose through evolution from a shorter gene composed of the kinase domain name. Further analysis revealed the presence of TTBK orthologues in genomes of sponges and cnidarians, indicating that this protein family is present throughout the animal kingdom (Physique 1B). Interestingly, TTBK orthologues were also found in slime molds (i.e., was present in the last common ancestor shared between animals and slime molds (Mycetozoa), but may have been lost during the evolution of fungi. Open.