Mycorrhizal fungi are mutualists that play crucial roles in nutrient acquisition in terrestrial ecosystems. Mycorrhizal symbioses arose repeatedly across multiple lineages of Mucoromycotina, Ascomycota, and Basidiomycota. Considerable variation exists in the capacity of mycorrhizal fungi to acquire carbon from soil organic matter. Here, we present a combined analysis of 135 fungal genomes from 73 saprotrophic, endophytic and pathogenic species, and 62 mycorrhizal species, including 29 new mycorrhizal genomes. This study samples ecologically dominant fungal guilds for which there were previously no symbiotic genomes available, including ectomycorrhizal Russulales, Thelephorales and Cantharellales. Our analyses show that transitions from saprotrophy to symbiosis involve (1) widespread losses of degrading enzymes acting on lignin and cellulose, (2) co-option of genes present in saprotrophic ancestors to fulfill new symbiotic functions, (3) diversification of novel, lineage-specific symbiosis-induced genes, (4) proliferation of transposable elements and (5) divergent genetic innovations underlying the convergent origins of the ectomycorrhizal guild.

The signal transduction pathway whereby the TxA2 (thromboxane A2) mimetic U-46619 activates vascular smooth muscle contraction was investigated in de-endothelialized rat caudal artery. U-46619-evoked contraction was inhibited by the TP receptor (TxA2 receptor) antagonist SQ-29548, the ROK (Rho-associated kinase) inhibitors Y-27632 and H-1152, the MLCK (myosin light-chain kinase) inhibitors ML-7, ML-9 and wortmannin, the voltagegated Ca2+-channel blocker nicardipine, and removal of extracellular Ca2+; the protein kinase C inhibitor GF109203x had no effect. U-46619 elicited Ca2+ sensitization in a-toxin-permeabilized tissue. U-46619 induced activation of the small GTPase RhoA, consistent with the involvement of ROK. Two downstream targets of ROK were investigated: CPI-17 [protein kinase C-potentiated inhibitory protein for PP1 (protein phosphatase type 1) of 17 kDa], a myosin light-chain phosphatase inhibitor, was not phosphorylated at the functional site (Thr-38); phosphorylation of MYPT1 (myosin-targeting subunit of myosin light-chain phosphatase) was significantly increased at Thr-855, but not Thr-697. U-46619-evoked contraction correlated with phosphorylation of the 20 kDa light chains of myosin. We conclude that: (i) U-46619 induces contraction via activation of the Ca2+/calmodulin/MLCK pathway and of the RhoA/ROK pathway; (ii) Thr-855 of MYPT1 is phosphorylated by ROK at rest and in response to U-46619 stimulation; (iii) Thr-697 of MYPT1 is phosphorylated by a kinase other than ROK under resting conditions, and is not increased in response to U-46619 treatment; and (iv) neither ROK nor protein kinase C phosphorylates CPI-17 in this vascular smooth muscle in response to U-46619.

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Clinical profiles differ between depressed children without and with sleep disturbance, with those presenting insomnia plus hypersomnia being most severely depressed. Differentiating depressed children with different sleep disturbances may have important implications for research efforts on the etiology and therapeutics of child depression.

A mechanism proposed for regulation of myosin phosphatase (MP) activity is phosphorylation of the myosin phosphatase target subunit (MYPT1). Integrin-linked kinase (ILK) is associated with the contractile machinery and can phosphorylate myosin at the myosin light-chain kinase sites. The possibility that ILK may also phosphorylate and regulate MP was investigated. ILK was associated with the MP holoenzyme, shown by Western blots and in-gel kinase assays. MYPT1 was phosphorylated by ILK and phosphorylation sites in the N- and C-terminal fragments of MYPT1 were detected. From sequence analyses, three sites were identified: a primary site at Thr(709), and two other sites at Thr(695) and Thr(495). One of the sites for cAMP-dependent protein kinase (PKA) was Ser(694). Assays with the catalytic subunit of type 1 phosphatase indicated that only the C-terminal fragment of MYPT1 phosphorylated by zipper-interacting protein kinase, and ILK inhibited activity. The phosphorylated N-terminal fragment activated phosphatase activity and phosphorylation by PKA was without effect. Using full-length MYPT1 constructs phosphorylated by various kinases it was shown that Rho kinase gave marked inhibition; ILK produced an intermediate level of inhibition, which was considerably reduced for the Thr(695)-->Ala mutant; and PKA had no effect. In summary, phosphorylation of the various sites indicated that Thr(695) was the major inhibitory site, Thr(709) had only a slight inhibitory effect and Ser(694) had no effect. The findings that ILK phosphorylated both MYPT1 and myosin and the association of ILK with MP suggest that ILK may influence cytoskeletal structure or function.

Although major depressive disorder (MDD) is associated with suicidal behaviors, some depressed individuals are not suicidal and others evidence various forms of suicidality. We thus investigated whether aspects of temperament and self-regulation of dysphoria represent risk factors for DSM-IV suicidality (recurrent thoughts of death, recurrent suicidal ideation, suicidal plan, and suicide attempt) in depressed youths. Using a sample of children with MDD (N = 407; ages 7-14 years), recruited from clinical sites across Hungary, we tested the hypotheses that: (a) suicidality is related to higher levels of trait negative emotionality as well as more maladaptive and fewer adaptive regulatory responses to dysphoria and (b) as the severity of suicidal behavior increases, levels of trait negative emotionality and dysfunctional emotion regulation also increase. We also explored if other aspects of temperament relate to suicidality. Children's DSM-IV diagnoses were based on semi-structured interviews and best-estimate psychiatric consensus. Parents independently provided ratings of their children's temperament, and children separately completed an inventory of emotion regulation (ER). Using multivariate models, we failed to confirm the hypothesized relations of negative trait emotionality and suicidality, but confirmed that high maladaptive and low adaptive ER response tendencies increase the odds of suicidal behaviors, above and beyond the risk posed by depressive illness severity. Unplanned interaction terms between temperament dimensions (other than negative emotionality) and ER suggested that at some high-extremes of temperament, ER has no impact on suicidality but in their absence, adaptive ER lowers the risk of suicidality. The practical implications of the findings are discussed.

During a clinical interview one must consider the age and sex of the child and the depressive state of the mother in assimilating information about the child.

The interactions of the catalytic subunit of type 1 protein phosphatase (PP1c) and the N-terminal half (residues 1±511) of myosin phosphatase target subunit 1 (MYPT1) were studied. Biotinylated MYPT1 derivatives were immobilized on streptavidin-biosensor chips, and binding parameters with PP1c were determined by surface plasmon resonance (SPR). The affinity of binding of PP1c was: MYPT1 1±296 . MYPT1 1±38 . MYPT1 23±38 . No binding was detected with MYPT1 1±34 , suggesting a critical role for residues 35±38, i.e. the PP1c binding motif. Binding of residues 1±22 was inferred from: a higher affinity binding to PP1c for MYPT1 1±38 compared to MYPT1 23±38 , as deduced from SPR kinetic data and ligand competition assays; and an activation of the myosin light chain phosphatase activity of PP1c by MYPT1 1±38 , but not by MYPT1 23±38 . Residues 40±296 (ankyrin repeats) in MYPT1 1±296 inhibited the phosphorylase phosphatase activity of PP1c (IC 50 = 0.2 nm), whereas MYPT1 1±38 , MYPT1 23±38 or MYPT1 1±34 were without effect. MYPT1 40±511 , which alone did not bind to PP1c, showed facilitated binding to the complexes of PP1c±MYPT1 1±38 and PP1c±MYPT1 23±38 . The inhibitory effect of MYPT1 40±511 on the phosphorylase phosphatase activity of PP1c also was increased in the presence of MYPT1 1±38 . The binding of MYPT1 304±511 to complexes of PP1c and MYPT1 1±38 , or MYPT1 1±296 , was detected by SPR. These results suggest that within the N-terminal half of MYPT1 there are at least four binding sites for PP1c. The essential interaction is with the PP1c-binding motif and the other interactions are facilitated in an ordered and cooperative manner.Keywords: myosin phosphatase; protein phosphatase-1; myosin phosphatase target subunit-1; surface plasmon resonance.Phosphorylation of the 20-kDa light chains of smooth muscle myosin (MLC20) on Ser19 plays a pivotal role in the regulation of contractile activity of smooth muscle and is also implicated in several functions of nonmuscle cells [1]. The phosphorylation level of myosin reflects the balance of activities of myosin light chain kinase and the myosin phosphatase (MP). Several different preparations of MP that are active with phosphorylated myosin and/or the phosphorylated form of MLC20 (P-MLC20) have been isolated from smooth muscle [2]. Recently several groups have suggested that the major form of MP in smooth muscle is a trimeric holoenzyme composed of the 38-kDa d isoform of type 1 protein phosphatase catalytic subunit (PP1cd), a large subunit of 110/133 kDa, and a small subunit of 20/21 kDa [3±6]. The 20/21 kDa subunit interacts with the large subunit and its function is not known [7,8]. The 110/133 kDa subunit binds to both the catalytic subunit of type 1 protein phosphatase (PP1c) and myosin [7±10] and thus is termed myosin phosphatase target subunit (MYPT) [11,12]. MP is not confined to smooth muscle. For example, PP1c complexed with a large subunit similar to MYPT that localized to the cytoskeletal and membrane fractions was identified in human platelets [13]. Recently,...