Lauryl Gallate Inhibits the Activity of Protein Tyrosine Kinase c-Src Purified from Human Platelets

The inhibitory effect of gallic acid (3,4,5-trihydroxybenzoic acid), and its ester derivatives methyl, propyl, octyl and lauryl has been tested on the tyrosine kinase activity of affinity purified c-Src from human platelets, using the artificial substrate Poly (Glu.Na, Tyr) 4:1. When tested as inhibitor of the autophosphorylation of the enzyme and the phosphorylation of the protein tyrosine phosphatase SHP-1 by c-Src, lauryl gallate was found to be a more potent inhibitor than other widely used protein tyrosine kinase (PTK) inhibitors such as genistein and herbimycin A. However, lauryl gallate did not inhibit the activity of the serine threonine kinases protein kinase A (PKA) and casein kinase II (CKII) from rat brain.     

Ultrastructure and some other properties of inverted thyroid follicles in suspension culture

Separated thyroid follicles in suspension culture invert in 5% serum. In some, the inversion is not complete in that a small normal follicle persists completely in the interior of an inverted follicle. In inverted follicles the lumens are distended and electron lucent. The bounding epithelial cells are stretched, have relatively few microvilli on the surface toward the medium but they have bundles of oriented microfilaments usually located near the lumen. The cells are connected together by tight junctions. When inverted follicles are punctured, the lumen shrinks, the cells retract and become cuboidal and microvilli reappear. Microfilaments persist at the luminal surface but no longer in oriented bundles. No appreciable extracellular matrix is present at the basal cell surface in contact with the lumen, but matrix is occasionally observed between cells. Since bundles of microfilaments like stress fibers are observed in the cells in suspension culture, the presence of stress fibers in cells in monolayer culture is probably not dependent on attachment but might be a reflection of the spreading of the attached cells.     

The TBC1D31/praja2 complex controls primary ciliogenesis through PKA‐directed OFD1 ubiquitylation

The primary cilium is a microtubule‐based sensory organelle that dynamically links signalling pathways to cell differentiation, growth, and development. Genetic defects of primary cilia are responsible for genetic disorders known as ciliopathies. Orofacial digital type I syndrome (OFDI) is an X‐linked congenital ciliopathy caused by mutations in the OFD1 gene and characterized by malformations of the face, oral cavity, digits and, in the majority of cases, polycystic kidney disease. OFD1 plays a key role in cilium biogenesis. However, the impact of signalling pathways and the role of the ubiquitin‐proteasome system (UPS) in the control of OFD1 stability remain unknown. Here, we identify a novel complex assembled at centrosomes by TBC1D31, including the E3 ubiquitin ligase praja2, protein kinase A (PKA), and OFD1. We show that TBC1D31 is essential for ciliogenesis. Mechanistically, upon G‐protein‐coupled receptor (GPCR)‐cAMP stimulation, PKA phosphorylates OFD1 at ser735, thus promoting OFD1 proteolysis through the praja2‐UPS circuitry. This pathway is essential for ciliogenesis. In addition, a non‐phosphorylatable OFD1 mutant dramatically affects cilium morphology and dynamics. Consistent with a role of the TBC1D31/praja2/OFD1 axis in ciliogenesis, alteration of this molecular network impairs ciliogenesis in vivo in Medaka fish, resulting in developmental defects. Our findings reveal a multifunctional transduction unit at the centrosome that links GPCR signalling to ubiquitylation and proteolysis of the ciliopathy protein OFD1, with important implications on cilium biology and development. Derangement of this control mechanism may underpin human genetic disorders.     

The avoidance of self-interference in the endemic daffodil Narcissus cyclamineus (Amaryllidaceae)

Hermaphrodite flowers usually possess floral traits to avoid the negative effects derived from inbreeding depression and/or self-interference between pollen export and reception, both acting as the main selective pressures on those floral traits. The avoidance of self-interference is widely accepted as the primary force promoting the separation between sexes within the flowers in time (dichogamy) and/or space (herkogamy) for self-incompatible species, which are already protected from the negative effects of inbreeding depression by the incompatibility system. Different degrees of incompatibility, herkogamy, and dichogamy have been reported for the genus Narcissus. However, the only mechanism for the separation of sexes reported up to date for Narcissus cyclamineus is herkogamy, while the presence of dichogamy and the type of incompatibility in this species remain uncertain. In this study, we analyze the patterns of sexual reproduction in N. cyclamineus to ascertain whether there is any selective pressure favouring sexual segregation or its maintenance and their mechanisms. N. cyclamineus is self-incompatible and dichogamy can be rejected for this species. Even though the species is self-incompatible, when cross-pollination is preceded by self-pollination the number of ovules available for legitimate crosses is diminished (ovule discounting). Pollinators are scarce during the flowering period, resulting in pollen limitation. It is suggested that both the scarcity of pollinators and ovule discounting may be acting synergically to promote herkogamy or its maintenance in this species.