Con taminating DNA was eliminated working with Turbo DNase, and RNA purity and concentration had been de termined working with a Nanodrop spectrophotometer. RNA quality was assessed working with an Agilent 2100 Bioanalyzer or by agarose gel electrophoresis just before RNA Seq or other applications, respectively. PCR amplification of a 196 nucleotide region of 16S rDNA was applied to con firm the absence of DNA before RNA Seq library creation. Library planning and RNA sequencing For each species, RNA samples from every single of three timepoints had been divided in two, with one particular half staying subjected to rRNA depletion employing the MICROBExpressTM Bacterial mRNA Enrichment Kit as per the guy ufacturers guidelines. Just about every from the rRNA depleted sam ples was then mixed to generate a species particular pool, such that each pool contained equal amounts of RNA from every time point.
Equivalent pools were produced to the untreated RNA samples. The two pools were destined to grow to be two separate li braries, the rRNA depleted pools have been sequenced using a protocol optimized for complete length transcripts, although the total RNA samples have been enriched for sRNAs selleckchem just before sequencing. All samples have been taken care of with tobacco acid pyro phosphatase to create five ends amenable for adapter ligation. To enrich for modest RNAs, the complete RNA sam ples were dimension chosen following polyacrylamide gel electrophoresis. RNA from the two li braries was then fragmented in the buffered zinc alternative and single stranded RNA adapters have been ligated towards the five and 3 ends to maintain strand specificity, just before re purifying on the polyacrylamide gel.
Every pool was then reverse transcribed and PCR amplified to make DNA colonies, which have been sequenced using an Illumina HiSeq 2000 sequencer. For your sRNA enriched library, go through lengths ranged from 24 to 94 nucleotides, while read through lengths had been as long as 150 nucleotides for your lengthy transcript enriched library. Alignment of reads to genomes Sequencing URB597 reads owning lower high-quality three ends have been trimmed utilizing the system PrinSeq. The high-quality trimmed reads have been aligned to their respective genome sequence utilizing Bowtie2 and then sorted, indexed and converted to BAM format employing SAMtools. For the complete length transcript libraries, 59,073,931, 44,462,362 and 94,358,187 reads were mapped towards the S. coelicolor, S. avermitilis, and S. venezuelae genomes respectively.
For your sRNA enriched libraries a complete of 21,871,239, 23,608,152 and 21,880,716 reads were mapped for the exact same respective genomes. The BAM files had been visualized applying Integrated Genomics Viewer. BEDTools was applied to calculate the optimistic and negative strand study depth at each and every nucleo tide, and also a Perl script was employed to exploit the BEDTools output in calculating the average sense and antisense coverage per nucleotide of each annotated gene. Non coding RNA analysis The genes with the highest levels of antisense coverage through the total length transcript libraries were selected for examination.

AUX1 encodes a membrane protein concerned in auxin uptake is really a crucial aspect for lateral root advancement and gravitropism in Arabidopsis. AUX1 like three unigenes have been down regulated in S2 and S3. Arabidopsis Aux/IAA proteins repressed transcription of auxin re sponsive reporter genes. The expression of 1 peanut AUX/IAA protein gene decreased in S2 to examine with S1. Two unigenes encoding AUX/IAA protein decreased drastically in S3 to evaluate with S1. These benefits have been coincided with all the loss of gravitropism after gynophore soil penetration. Together, our results indicated that auxin biosynthesis, transportation and signaling may signify key downstream effectors of light signaling during gynophore growth and pod initiation.
On top of that, selleck chemicals genes in gibberellins, ABA and ethylene biosynthesis or signaling have been detected to up or down regulated in these three stages. In recent years, considerable research have revealed the important thing regulatory roles of brassinosteroid in plant development and development. Brassinosteroid can be perceived by BRI1 and after that activates BZR family members of transcription factor which could physically interact with PIFs and DELLA protein. Consequently, modify in biosynthesis or signaling of BR would not only affect endogenous hormone degree and/or hormone signalling but would also have an impact on light signals and responses. Nonetheless, there’s no report about BR written content or signal transduction on peanut pod create ment. Interestingly, we observed in S2 the decreased ex pression of 1 brassinosteroid receptor and 3 BRASSINOSTEROID INSENSITIVE 1 related recep tor kinase unigenes.
In S3, beside three BRI1 associated receptor kinase 1 unigenes, 1 brassinosteroid LRR receptor kinase unigene and two brassinosteroid bio synthetic protein unigenes were down regulated. Two brassinosteroid receptor unigenes IPI-145 1201438-56-3 have been up regulated in S3. These altered expression patterns of BR biosynthesis and signal transduction connected genes recommended the involvement of brassinosteroid in gynophore elongation and pod enlargement initiation. On alterations in light signaling, improvements in hormone biosynthesis/signaling, down stream alteration related to cell division and cell wall formation were observed within this examine. Cellulose is definitely the significant part of plant cell wall and cellulose synthase catalyse the for mation of cellulose.
Mutation of cellulose synthase gene triggered lessen in cellulose material and resulted in cell swelling, restrict root and hypocotyl elongation. Thirty eight unigenes encoding cellulose synthase had been down regulated in S3 in contrast to S1, indicating a drastic change in synthesis of cell wall parts below dark problem. Other genes involved in cell di vision, cell cycle and cell elements are substantially enriched among unigenes with 2 folds changes in S3.

Specific ABC proteins are drug efflux trans porters found inside the cell membrane, and also have roles while in the biochemical defence towards toxicants. The ABC transporter P glycoprotein transports IVM and has relevance like a biochemical factor limiting the medicines toxicity in mice and nematodes. It’s been suggested that P glycoprotein can be implicated within the resistance of pests and parasites to AVMs, plus a salmon louse homologue of P glycoprotein identified as SL PGY1 has become cloned. During the present examine, mRNA expression of SL PGY1 was unaffected by EMB deal with ment and did not differ involving S and PT strains, confirming very similar data previously reported for these strains. Furthermore to comparing transcriptional profiles be tween louse strains from the absence of drug publicity, the existing examine also addresses transcriptomic responses of the two louse strains to brief phrase exposures to 200 ug L one of EMB.
The selected EMB concentration is just not toxic for the resistant salmon louse strain, but during the drug vulnerable strain it ends in near to 100% immotility after 24 hrs exposure. Following one to three hours of publicity, handful of transcriptomic responses have been observed in the resistant strain, and thus these information tend not to give evidence for the presence of certain resistance mechanisms requiring selleck peptide company transcriptional regulation. In the drug susceptible strain, a complicated array of transcriptional responses to EMB was observed. This locating was anticipated, as this level of EMB publicity was acknowledged to lead to sizeable toxic responses just after 24 hours.
Early transcriptional responses are therefore likely to comprise each adaptive and common pressure responses, likewise as tran scriptional improvements reflecting the toxic action Ginkgolide B from the drug. Conclusions The transcriptional profiling of a drug susceptible and an EMB resistant laboratory strain of salmon lice re vealed diminished mRNA expression of the number of LGIC subunits inside the EMB resistant lice. GluCl, considered to be the key target web-site of AVM medication in invertebrates, displayed similar mRNA expression levels in EMB resistant and reference strains. In contrast, subunits of GABA Cl and nAChR showed decreased mRNA abun dances in the EMB resistant compared to the reference strain. When GABA Cl is viewed as a secondary target of AVMs in invertebrates, nAChR will not be historically thought of a target internet site for AVMs, while it has been shown to interact with AVMs in vertebrates.
It is actually achievable that nAChR and GABA Cl signify added EMB target websites in salmon lice, and that the down regulation of those channel subunits within this EMB resistant strain could be linked towards the resistance pheno sort. In the present review no adjustments were observed from the expression levels of biotransformation enzymes and drug transporters, the two of which classes are advised to contribute to AVM resistance in other species.

This server shall be practical to narrow down the time and cost required to display a chemical library. Background Tuberculosis caused by Mycobacterium tuberculo sis, is among the main leads to of morbidity and mortal ity inside the producing world. Tuberculosis manifests in lots of clinical types from an energetic sickness state to clini cal latency which can extend for decades. It has been reported that there have already been 9. four million new situations of TB with an estimated international mortality of 1. 7 million within the year 2009. Recent reviews propose that a high proportion, averaging about 85% situations are actually accounted to come about in Asia and Africa with India and China alone accounting to 50% from the total burden of illness. Evidences also point to an raising inci dence of drug resistant TB in excess of the previous decade. Furthermore, its catastrophic synergy with Acquired Immune Deficiency Syndrome have made it a major overall health concern worldwide.
The present medication used in the first a total noob line therapy of tuberculosis has become discov ered no less than half a century ago, as well as the unabated worldwide rise of tuberculosis calls for your advancement of novel tools and solutions for fast and effective identification of novel molecules with anti tubercular activities. Using the lack of complete techniques level beneath standing in the causative organism and its intricate bio logical pathways and handle mechanisms, it’s been recommended that total cell phenotypic screens offer a bet ter proposition in comparison with single gene primarily based biological screens. The availability of approaches for large throughput screening has drastically contributed for many such datasets staying created obtainable in public domain. Using the pretty very low hit fee for such higher throughput biological screens, it has turn out to be inevitable to prioritize molecules to become taken up for biological screens.
It has been suggested out that virtual screening of huge compound libraries selleck working with computational meth ods like machine studying methods could possibly be efficiently employed as being a complementary approach to phenotypic screens in drug discovery. The availability of smaller molecule bio assay datasets in public domain pro vide a precious suggests to construct predictive computational models which will be possibly utilized to prioritize mole cules for biological assays from massive digital databases. We now have previously employed machine understanding approaches to classify inhibitors identified from bioassay screens of Mycobacterium tuberculosis in Middlebrook 7H12 broth. The 7H12 media enriched Middlebrook 7H9 broth allows speedy recovery of mycobacteria from clini cal specimens, sputum, and respiratory secretions. Though the ADC enrichment provides additional sensitivity to your microbial culture, albumin acts like a protective agent by binding free fatty acids, which could be toxic to mycobac terium species and catalase destroys toxic peroxides that could be current inside the medium and dextrose acts as an energy supply.

To date, bacteriocins have only been recognized in Prochlorococcus marinus MIT9313. The toxin microcystin synthetase gene cluster was shown to become of ancient origin. Earlier envir onmental research showed the steady presence of mutant cyanobacterial mcy gene clusters, which were inactivated both in freshwater samples by ISs and in brackish water by MITEs. Note that the intact hassallidin gene cluster continues to be retained in parallel cultures of mutant Anabaena sp. 90, which has an inactivated anabaenopep also demonstrated the incidence of genes inactivated by IS transposition, such as some with very important functions, within the population of Synechococcus strains in scorching spring mats. Higher frequencies of mobile genetic element derived pseudogenes are more likely to be frequent between transposable component wealthy cyanobacteria, espe cially in strains of bloom forming genera, but as however re key undocumented.
Anabaena sp. 90 dedicates 5% of its genome to bio synthesis of modest peptides, such as hepatotoxic micro cystins along with the protease inhibitors anabaenopeptins and anabaenopeptilides. Involving 3% and selleck chemical 4% on the genomes is generally devoted towards the biosynthesis of secondary metabolites between sequenced cyanobacterial genomes. Even so, former works only counted non ribosomal gene clusters and missed the ribosomal bio synthesis pathways. Here, we took into account each ribosomal and nonribosomal gene clusters. Considering the widespread occurrences of ribosomal pathways in cyanobacteria and that nonribosomal gene clus ters also amount to three. 5% of Anabaena sp. 90 genome, it appears that about 5% in the genome is typically assigned to normal merchandise biosynthesis in cyanobacteria. MK-0752 An additional nonribosomal gene cluster responsible for manufacturing of the antifungal compounds hassallidins was identified throughout genome assembly, primarily be lead to the gene cluster was inactivated by a deletion occasion.
Also, the genome sequence led us to dis cover the ribosomal production pathway for cyanobac tins and their frequent occurrence in Anabaena as tilide synthetase gene cluster. Our outcomes showed that people mutants with an inactive hassallidin biosyn thetic pathway prevailed over cells with functional genes during the culture. This might indicate a growth benefit for cells with mutated bioactive compound synthetase gene clusters beneath laboratory ailments, possibly resulting from a decrease metabolic burden. Similarly, we detected a loss of gasoline vesicles, the sub cellular structures responsible for buoyancy, which are crucial for bloom forming planktonic cyanobacteria. This phenotype is derived in the truncated gvpG gene while in the gvp operon. The truncation most prob ably was picked once the strain was purified on sound media.

Interestingly, in yet another genus on the Actinomycetales with circular genomes, Corynebacterium, this positional bias is simply not observed. This raised the question irrespective of whether this degree of conser vation and synteny is far more pronounced if numerous species are taken into consideration, i. e. for your genes producing up the core genome from the family. As a to start with step to response this question, the family members core genome was calculated, when once more applying EDGAR. Inside a to begin with stage the core genome was calculated for every feasible exceptional subset on the seven genomes. Following this, an exponential decay perform was fitted on the observed core genome counts for every genome quantity as described by Tettelin et al. Utilizing this perform, the improvement within the core given a presumed sequential addition of a lot more genomes was extrapolated.
This led to the predicted core genome of about 810 genes, i. e. the theoretical core of all Pseudonocardiaceae. Using the core genome of these seven species comprised of 864 genes the 810 worth is almost reached using the seven genomes applied. A check in cluding two Streptomyces species only reduced the amount of genes during the resulting core by about 45 genes to 765 which is by now reached with the selleck PF299804 then nine genomes. That is pretty surprising, as Streptomyces species often possess linear ends. This implies that Pseudonocardiaceae and Streptomycetaceae, despite getting various chromosome topologies, retain their core genes near to the oriC. This implication is backed by further analyzing the correlation among gene conservation and spot rela tive towards the oriC, the core genes are found predominantly clustered all-around the oriC even though genes conserved in only some species plus the two,967 S.
espanaensis singletons are observed farther far from the oriC. The genes on the accessory genome of S. espanaensis are either ancient and/or obtained from closely connected species The distribution of core and accessory genes in the gen ome raises the question of what mechanism is creating selelck kinase inhibitor this peculiar pattern. As many independent circularization occasions are exceedingly unlikely, the genome of your com mon ancestor on the Pseudonocardiaceae was either incredibly big and suffered gene reduction for the duration of speciation or it had been rather minor and a lot of the accessory genes had been acquired later on. This in turn raises the question of irrespective of whether there is certainly evidence the accessory genes were acquired lately.
To shed light on this timing, we carried out a principle element examination in the relative polynucleotide frequencies of all the genes. Examination of polynucleotide frequencies had been proven for being practical for classification of quick DNA fragments obtained from distinct genomes. In our situation, no substantial differences for the dinucleotide level may be observed for many from the genes, neither when comparing core and accessory genes, nor when comparing genes near and distant for the oriC.

Since the NRPS portion in every single from the five PKS NRPS hybrids in G. lozoyensis genome is made up of just one A T C module, one amino acid is predicted to become added to your polyketide made through the PKS portion within the cluster, much like ApdA in a. nidulans and ATEG00325 inside a. terreus, which are involved from the biosynthesis of aspyridones and flavipucine, respectively. Comparing the rich genetic possible for secondary me tabolites from the G. lozoyensis genome, only pneumocandins had been previously identified in the fungus. In an try to search out more chemistries, we recognized isolecanoric acid and pseudogyrophoric acid as two new fermentation goods of G. lozoyensis. For that reason, bulk of your secondary metabolites in G. lozoyensis continue to be to become characterized.
Despite the advances during the discipline of microbial secondary metabolite biosynthesis, how the fundamental biology, ecology, and trophic tactics of microorganisms relate to their secondary robertsii by genomic sequencing. Genomic mining of a few Aspergillus spp. has led to the elucidation of biosyn thetic this content pathways of a variety of bioactive compounds, like terrequinone A, emericellamide, aspyridones, pyripyropene A and echinocandin B. Genomic sequence examination showed that G. lozoyensis has the likely to provide a varied array of all-natural goods. The genome was predicted to encode 49 gene clusters that contribute to its secondary metabolome, metabolite manufacturing remains poorly understood. Application of effective methods to mine the metabolite encoding gene clusters in G.
lozoyensis and also other poorly acknowledged fungi, though identifying their CI1040 corresponding metabolites, presents a challenge and chance for natural goods discovery. GLNRPS4 and GLPKS4 are centrally located during the pneumocandin biosynthetic gene cluster, and the way they cooperate with other genes while in the cluster is still speculative. Despite the fact that they can be independently tran scribed and translated, their transcription is more likely to be synchronized or co regulated. The 1st module in GLNRPS4 features a one of a kind T C A T C structure, and also the to start with T domain during the T C A T C module is suggest to accept thiolated intermediates as uncovered in emericellamide biosynthesis or adenylated substrates similar to yersiniabactin biosynthesis. Consequently, the first T domain while in the T C A T C module of GLNRPS4 could be respon sible for accepting the incoming ten,twelve dimethylmyristoyl side chain intermediate, whereas the 2nd T domain would accept the four,five dihydroxyornithine adenylated by the modules A domain. Threonine, four hydroxyproline, 4,5 dihydroxyhomotyrosine, 3 hydroxyglutamine and 3 hydroxyproline/3 hydroxy four methylproline might be sequentially added to the expanding chain steady with all the in silico prediction that the A3 in GLNRPS4 is particular to proline.

Cytological studies have established that female Lepidoptera may perhaps kind syn aptonemal complexes in early meiotic prophase I, but no recombination nodules are formed subsequently. Alternatively, a construction known as elimination chromatin is formed. Typically chiasmata are formed from retained pieces on the SC in which a RN is, or is, present. The formation on the chiasmata will take place during the cell destined to come to be the oocyte from the D. melanogaster germarium. Four genes appear crucial in D. melanogaster for SC formation and thus probably chiasmata formation, cross more than suppressor on 2 of Manheim M, crossover sup pressor on three of Gowen G, corona and nipped B. No genes unique for RN alone can be identified on FlyBase. Pararge aegeria females only express nipped B, which can be concerned in a variety of cel lular processes in D.
melanogaster such as mitosis. It truly is also the only among the 4 SC genes for which orthologs outdoors Drosophila selleck chemical natural product library might be identified. Ra ther interestingly, a large proportion of the genes in volved in D. melanogaster meiotic chromosome cohesion and segregation also appeared to be Drosophila or Dip tera particular and weren’t recognized during the P. aegeria transcriptome. These incorporate grauzone, corona, orientation disrupter and mei S332. A number of genes are, on the other hand, highly conserved and orthologs have been discovered in Lepidoptera as males do display crossing above. These include each mei W68 and mei 218 but in particu lar consists of the important meiotic checkpoint gene pch2. Female P. aegeria did not express any of these genes. The P.
aegeria oogenesis transcriptome described here is thus in accordance selleck using the preceding observations made throughout cytological scientific studies on female Lepidoptera. Vitellogenesis and lipid storage Not merely is cell cycle regulation coordinated with oocyte differentiation in D. melanogaster, but also with re supply provisioning of your oocyte. The gene greatwall, for instance, is the two essential in D. melanogaster for maternal provisioning of your egg all through vitellogenesis and to make sure secondary meiotic arrest by stage 14 of oogen esis in metaphase I. It’s a highly conserved gene in Metazoa and P. aegeria females did express this gene in the course of oogenesis. Fur thermore, gwl interacts with polo kinase in mitotic regulation especially through early em bryogenesis, and is maternally offered. Transcripts of the two have been detected in P. aegeria oocytes. Vitellogenesis all through insect oogenesis is characterised by the accumulation within the establishing oocytes of substantial lipid transfer proteins, this kind of as Vitellogenin and Apolipophorins. Predominantly, LLTPs are produced in the excess fat bodies and secreted to the hemolymph, but not all yolk proteins are extraovarian.

Prior research also demonstrated that aromatase exercise from the endometrium plays a crucial part during the malignant transformation of endometrial cells by converting androgen into mitogenic estrogen from the endometrial tissue. To determine the position of aro matase in non genomic signaling pathway mediated by testosterone, we examined testosterone stimulated ERK and Akt phosphorylation in Hec1A cells pre treated by letrozole, an aromatase inhibitor. As expected, letrozole abrogated the phosphorylation of ERK and Akt stimulated by testosterone. Additionally, we also identified that letrozole therapy lowered expression levels of aromatase in Hec1A cells. These data strongly recommend that aromatase is concerned in testosterone actions in cells express ER 36.
Discussion Estrogen receptor is a member from the nuclear receptor superfamily and perform as ligand dependent transcrip tion factor inside the nucleus to mediate estrogen signaling. Nevertheless, accumulating evidence show that there’s a quick estrogen signaling which can’t be explained by genomic signaling pathway that ordinarily will take hrs to perform. Lately, we selleck chemicals located that ER 36 was expressed in ER constructive and ER detrimental breast cancer cells, suggesting that ER 36 expression is regulated in a different way from ER 66. During the existing research, we located that ER 36 is expressed primarily on the plasma membrane in ER 66 adverse endome trial cancer Hec1A cells and ER 36 mediates membrane initiated MAPK ERK and PI3K Akt pathways induced by testosterone. It’s been reported that endometrial cancer danger is enhanced in the two pre and postmenopausal women with elevated plasma levels of testosterone.
Early from the neoplastic system, abnormal endometrial cells can locally make estrogens in the plasma pool of andro gen, and so obtain a growth benefit independent of cir culating estrogens. The Flavopiridol regional concentration of estrogens in endometrial cancer was reported to get larger than that from the blood and also the endometrium of cancer absolutely free gals. Without a doubt, prior studies have proven that aromatase exercise is improved in endometrial cancer cells, but not regular endometrial cells. Furthermore, elevated circulating androgen has also been related with hyperplasia with the endometrium, which generally precedes and accompanies the occurrence of sort I endometrial carcinomas. Aromatase is really a crucial enzyme during the synthesis of estrogen that is definitely responsible for binding of testosterone and catalyzes the series of reactions even tually resulting in estrogen manufacturing. Earlier reports demonstrated that aromatase is current in endometrial cancer tissue, suggesting that aromatase plays a part in converting testosterone into mitogenic estrogens in endometrial tissue.

The African Swine Fever Virus protein A238L incorporates a very similar motif the PKIIITG motif. LxVP peptides are derived from the conserved cal cineurin docking NFATc motif LxVP and compete with NFATc to the binding to activated calcineurin. The NFATc isoforms differ from the affinity of their LxVP motifs towards calcineurin. Pep3 is derived from your CNBR2 of NFATc3. This sixteen amino acid oli gopeptide contains the LxVP motif, binds to purified and cellular calcineurin and competes with GST CNBR2 for binding to calcineurin. Retroviral overexpression of Flag Pep3 within the murine D10G4. 1 TH2 cell line impaired the expression of IL five, IL six and IL 13 along with the nuclear translocation of NFATc3 soon after PMA cal cium ionophore stimulation. NFATc3, but not NFATc2 and NFBactivation is impacted by Pep3. The LxVPc1 peptide, spanning the 15 amino acids of human NFATc1 371 385, disrupts calcineurin NFATc1 and c2 binding.
GST LxVPc1 binds to calcineurin a lot more efficiently than any in the PxIxIT motifs of NFATc1 to c4. The GST 2-ME2 molecular weight LxVPc2 fusion peptide from NFATc2 was not able to bind to calcineurin below exactly the same problems. The LxVPc1 peptide inhibits calcineurin phosphatase action about the RII phosphopeptide and increases the phosphatase action on pNPP. Overexpression of GFP LxVPc1 fusion protein in HeLa cells inhibits NFATc2 dephosphorylation and nuclear translocation on ionophore remedy.in Jurkat T cells it inhibits NFATc2 dephosphorylation and the expression of luciferase underneath handle in the IL two or RCAN1 four promoter on PMA ionophore stimulation. Protein fragments based upon other motifs have been derived from CABIN1 and RCAN1. The protein fragment CABIN1700 901 inhibits the dephosphorylation on the RII phosphopeptide by calcineurin within a noncompetitive man ner.
Overexpression of CABIN1700 901 in HEK293 cells coexpressing constitutively active calcineurin inhibits the dephosphorylation of NFATc2, its nuclear translocation selleck chemical and luciferase reporter gene expression beneath NFAT con trol. Overexpression of this fragment in Jurkat T cells sup presses the expression of luciferase controlled from the IL two promoter upon PMA ionomycin stimulation. RCAN1 and two include a SP repeat motif binding for the cat alytic centre of calcineurin. The SP repeat peptide, which could be phosphorylated by MAPK and GSK three, simulates a substrate for calcineurin and therefore inhibits calcineurin action towards RII phos phopeptide in the competitive method in cell free assays. This inhibitory effect is independent from the phosphoryla tion status in the peptide. On the other hand, overexpressed RCAN1 fragments containing only the SP repeat domain do not suppress calcineurin NFATc signalling in cells. A peptide containing the CIC motif and also the C terminal 30 amino acids of RCAN1 blocks dephosphor ylation on the RII phosphopeptide by calcineurin, but nei ther the CIC containing peptide nor the C terminus alone.