MassIVEapplication/xmlftp://massive-ftp.ucsd.edu/v05/MSV000090945/primaryOK2000000Justyna Labuzhttps://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=e10bc6a8fe614e1ca3f179e01af99cffjustyna.sojka@uj.edu.plMassIVE26UNIMOD:4 - "Iodoacetamide derivative."UNIMOD:1 - "Acetylation."UNIMOD:35 - "Oxidation or Hydroxylation."ProteomicsQ ExactiveArabidopsis Thaliana (ncbitaxon:3702)Malopolska Centre of Biotechnology, Jagiellonian University<h4>Background</h4>This study examines the biological implications of an overlap between two sequences in the Arabidopsis genome, the 3'UTR of the PHOT2 gene and a putative AT5G58150 gene, encoded on the complementary strand. AT5G58150 is a probably inactive protein kinase that belongs to the transmembrane, leucine-rich repeat receptor-like kinase family. Phot2 is a membrane-bound UV/blue light photoreceptor kinase. Thus, both proteins share their cellular localization, on top of the proximity of their loci.<h4>Results</h4>The extent of the overlap between 3'UTR regions of AT5G58150 and PHOT2 was found to be 66 bp, using RACE PCR. Both the at5g58150 T-DNA SALK_093781C (with insertion in the promoter region) and 35S::AT5G58150-GFP lines overexpress the AT5G58150 gene. A detailed analysis did not reveal any substantial impact of PHOT2 or AT5G58150 on their mutual expression levels in different light and osmotic stress conditions. AT5G58150 is a plasma membrane protein, with no apparent kinase activity, as tested on several potential substrates. It appears not to form homodimers and it does not interact with PHOT2. Lines that overexpress AT5G58150 exhibit a greater reduction in lateral root density due to salt and osmotic stress than wild-type plants, which suggests that AT5G58150 may participate in root elongation and formation of lateral roots. In line with this, mass spectrometry analysis identified proteins with ATPase activity, which are involved in proton transport and cell elongation, as putative interactors of AT5G58150. Membrane kinases, including other members of the LRR RLK family and BSK kinases (positive regulators of brassinosteroid signalling), can also act as partners for AT5G58150.<h4>Conclusions</h4>AT5G58150 is a membrane protein that does not exhibit measurable kinase activity, but is involved in signalling through interactions with other proteins. Based on the interactome and root architecture analysis, AT5G58150 may be involved in plant response to salt and osmotic stress and the formation of roots in Arabidopsis.Phototropin2 3'UTR overlaps with the AT5G58150 gene encoding an inactive RLK kinase.Łabuz Justyna J, Banaś Agnieszka Katarzyna AK, Zgłobicki Piotr P, Bażant Aneta A, Sztatelman Olga O, Giza Aleksandra A, Lasok Hanna H, Prochwicz Aneta A, Kozłowska-Mroczek Anna A, Jankowska Urszula U, Hermanowicz Paweł PF13M22.21, Transphosphorylases, Materials, Genetic, PSCTK5, Btkl, RLK, Rlk, K3G17.4, UTR, Gene, INSDC_feature:gene, UTR2, Transphosphorylase, Cistrons, Phosphotransferases, MCK7.2, receptor lectin kinase, K3G17_4, GPR14, UR-2-R, receptor-like kinase, PTK4, untranslated region, Material, BTKL, MCK7_2, Gpr14, A130089B16Rik, Genetic Materials, F13M22_21, Cistron, TKL, Kinase, Kinases., ATP Phosphotransferases, Phosphotransferase, Genetic Material, PTK-RL-18, ATPNetworks, hydrogen transport, Materials, Kinship, SAPKa, determination, 2410041A17Rik, ACTG, A., ACTE, postnatal development, adenosinetriphosphatase activity, Visible Light, growth and development, fs(1)M34, GRO:0005338, BSK, Bsk, csp2, Act5, Cardaminopsis, L-Leucine, l(1)G0420, DmelCG12051, Membrane Tissues, L-Isomer Leucine, intracellular localization, primordia, Arabidopsis thalianas, ATP hydrolysis-dependent activity, A. thalianas, hnu, CG9063, symptoms, A130089B16Rik, HEL-176, Life Cycle, Brassinosteroid, CG4601, Kinase, phosphokinase activity, Leucin, foton, bsk, PTK-RL-18, Family Life Cycle, multicellular organismal biosynthetic process, imprinted and ancient gene protein, A, single organism cellular localization, C, single-organism biosynthetic process, (Ca2+ + Mg2+)-ATPase, cyt5C, Kinship Network, Genomes, DBSK/JNK, DHO, Junk, L, anabolism, Mouse-ear Cress, membrane region, Tissue, hypoplasia, CG18572, localization within cell, BcDNA:GH03694, Mouse-ear, TNFSF14, phototropin 2, PTK4, climbing root (narrow), act 42A, signaling process, Ac5C, bud, CG4027, c-Jun, ACT, Act, NON PHOTOTROPIC HYPOCOTYL 1-LIKE, Family Life Cycles, ATP Phosphotransferases, Arabidopses, DmelCG9063, ATP, gamma, single organism signaling, homodimer, Membrane Tissue, screening, NPL1, hydrogen ion transmembrane transport, D-junk, Radiation, l(1)G0330, ATP hydrolase activity, hydrogen ion transport, Actin/BAP47, UNQ391/PRO726, dJNK, CTE-II, AACT, K3G17.4, BSK/IYK, CTE-IIa, integral to membrane, aerial root (narrow), act, Ach1, hBACH, ACH1, A. thaliana, Light, future organ, membranous organ component, Rek7, LACH1, act42A, K3G17_4, GIG25, LIGHT, down the electrochemical gradient, ATPase-dependent activity, Su(b), Alpha-1-antichymotrypsin His-Pro-less, AFFX-Dros-ACTIN_M_r_at, GIG24, DFNA26, Genetic Materials, actin, overlap, TKL, Filiation, LACH, GAT, DFNA20, Genetic Material, cellular localisation, Visible Radiations, ATP phosphohydrolase activity, ATP hydrolysis coupled proton transport, Transphosphorylases, Visible Radiation, membrane of organ, DmelCG4027, HVEML, PSCTK5, BcDNAGH03694, l(1)G0117, signs, SV40 T-antigen, Leuzin, Actin, INSDC_feature:gene, whole genome, root, establishment and maintenance of cellular localization, beta-actin/Bap47, 3L6, l(1)G0486, Life Cycles, l(1)G0245, DJNK, SNF1-RELATED PROTEIN KINASE 2-2, Arabidopsis, actin5C, light quantum, l(1)G0009, rich, Material, SNF1-related protein kinase 2.2, ACTL3, Lach1, Cistron, Hleu, E430016J11Rik, Protein Kinase, BACH, BSK/DJNK, GTK, l(1)Ab, Act5c, (RS)-Leucine, Els1, biological signaling, l(1)G0010, photoreceptor, Family Member, Ehk1, establishment and maintenance of localization in cell or cell membrane, L Isomer, ACT5C, Gene, CG12051, Network, biosynthesis, Serpin A3, Ximpact, IPOA, Bach, Ly113, l(1)G0025, receptor-like kinase, reduced, Hek7, integral component of membrane, ATP monophosphatase activity, BTKL, thalianas, Gene Products, AW125296, Cek7, Cell growth-inhibiting gene 24|25 protein, tiny, Phosphotransferase, adenosine triphosphatase activity, DmelCG5680, DJNK/bsk, study, proton transport, act5C, complex V (mitochondrial electron transport), Genetic, single-organism cellular localization, Cresses, Research, formation, Tissues, Btkl, Lichtquant, BRWS2, cell elongation, RLK, Rlk, ATPase activity, Transphosphorylase, Visible, (+-)-Leucine, ATP-protein phosphotransferase, synthesis, fs(1)829, K21L19.6, AI854630, Act42a, anon-EST:fe2D2, SNRK2-2, 1700027G07Rik, SOD, DmelCG18572, region of membrane, photon, K21L19_6, Cress, TR2, T11, Mouse ear, Kinship Networks, rudiment, Family, Act-5C, Cte-II, small, l(1)G0177, F13M22.21, membrane, 42A, T DNA, Jnk, JNK, findings, CPS, MCK7.2., ALS, Family Research, adenosine 5'-triphosphatase activity, Arabidopsis thaliana, Proteins, dJ393D12.2, L-Isomer, Cistrons, CD258, Phosphotransferases, impact-a, MCK7.2, all, Family Members, development, JNK/SAPK, SRK2D, jnk, CAD, passive proton transport, beta-actin, MCK7_2, D-JNK, Protein, chemical analysis, HCO3--ATPase, M32055, whole membrane, IMPACT, imprinted and ancient gene protein homolog, F13M22_21, C85044, Act42, ALS1, ACTA3, coupled, Radiations, l(1)G0079, transmembrane, SPK-2-2, hydrogen transmembrane transport, underdeveloped, Leucine, Kinases, HVEM-L, PYR1, VSCM, Photoradiation, hSod1, postnatal growth, act 5C, thaliana, placode, Membrane, Mouse-ear Cresses, LTg, receptor lectin kinase, signalling, Protein Gene Products, Gene Proteins, Photoradiations, signalling process, Families, HEL-S-44, CG5680, DL-Leucine, 2-amino-4-methylpentanoic acid, DRORUD, BAP47, assay, ACTSG, PROTEIN KINASE, Relatives, growth, Bap47, RWDD5, Leu, RAK, Actin5CF13M22.21, Materials, Genetic, PSCTK5, Btkl, RLK, Rlk, receptor lectin kinase., K3G17.4, UTR, Gene, INSDC_feature:gene, UTR2, Cistrons, MCK7.2, K3G17_4, GPR14, UR-2-R, receptor-like kinase, PTK4, untranslated region, Material, BTKL, MCK7_2, Gpr14, A130089B16Rik, Genetic Materials, F13M22_21, Cistron, TKL, Genetic Material, PTK-RL-18extent, Forms, hydrogen transport, Stocks, Materials, Kinship, SAPKa, 2410041A17Rik, A., response to salinity, A4, adenosinetriphosphatase activity, Visible Light, fs(1)M34, Cardaminopsis, DmelCG12051, L-Isomer Leucine, intracellular localization, protein polypeptide chains, A. thalianas, hnu, ionic compounds, CG9063, A130089B16Rik, HEL-176, Life Cycle, Msal-1, Brassinosteroid, CG4601, Kinase, Polymerase Chain, phosphokinase activity, PTK-RL-18, imprinted and ancient gene protein, A, gamma sarcoglycan, C, cyt5C, Kinship Network, Genomes, DBSK/JNK, DHO, Junk, L, anabolism, Tissue, Salt, CG18572, Msal, proteins, BcDNA:GH03694, sel, Salz, Mouse-ear, salts, GPR14, ACE, salt, Blue, climbing root (narrow), Gpr14, ACT, Act, ATP Phosphotransferases, xCYP26, single organism signaling, hydrogen ion transmembrane transport, Anchored Polymerase Chain Reaction, Radiation, completeness, Actin/BAP47, UNQ391/PRO726, dJNK, CTE-II, land plants, AACT, K3G17.4, Racial, CTE-IIa, integral to membrane, aerial root (narrow), act, Ach1, hBACH, ACH1, Light, act42A, GIG25, common salt, LIGHT, down the electrochemical gradient, Su(b), AFFX-Dros-ACTIN_M_r_at, GIG24, actin, sarcoglycan, TKL, LACH, PCR, ATP phosphohydrolase activity, DmelCG4027, HVEML, PSCTK5, ionic compound, BcDNAGH03694, SV40 T-antigen, Leuzin, INSDC_feature:gene, whole genome, root, UTR2, 3L6, polymerase chain reaction, DJNK, higher plants, actin5C, light quantum, rich, Material, SNF1-related protein kinase 2.2, 35kD dystrophin-associated glycoprotein, Hleu, Continental Population, E430016J11Rik, BACH, GTK, l(1)Ab, Act5c, (RS)-Leucine, Els1, photoreceptor, Ehk1, ACT5C, Polymerase Chain Reactions, biosynthesis, protein-containing complex, Bach, Ly113, receptor-like kinase, ra&#237z lateral (Spanish, ATP monophosphatase activity, BTKL, thalianas, Gene Products, Cek7, Cell growth-inhibiting gene 24|25 protein, Continental Population Group, MAM, gamma-SG, adenosine triphosphatase activity, DmelCG5680, DJNK/bsk, plantae, anon-EST:fe3D10, proton transport, Cresses, Tissues, 側根 (Japanese, Btkl, DmelCG8827, Lichtquant, BRWS2, RLK, Rlk, ANCE, (+-)-Leucine, synthesis, fs(1)829, K21L19.6, ance, anon-EST:fe2D2, DmelCG18572, Kochsalz, AnCE, photon, Cress, Anchored PCR, TR2, T11, Act-5C, 42A, CPS, Family Research, 35 kDa dystrophin-associated glycoprotein, Arabidopsis thaliana, Proteins, XCYP26A1, dJ393D12.2, UTR, L-Isomer, CD258, MCK7.2, SGCG, all, Family Members, JNK/SAPK, SRK2D, native protein, Natriumchlorid, passive proton transport, beta-actin, D-JNK, chemical analysis, HCO3--ATPase, M32055, IMPACT, imprinted and ancient gene protein homolog, C85044, Act42, background, NaCl, INSDC_feature:regulatory, coupled, DMDA1, HVEM-L, PYR1, thaliana, B130022O04Rik, primary structure of sequence macromolecule, receptor lectin kinase, signalling, Gene Proteins, Photoradiations, signalling process, Families, Dietary Sodium, CG5680, DL-Leucine, 2-amino-4-methylpentanoic acid, DRORUD, BAP47, ACTSG, PROTEIN KINASE, Relatives, Bap47, Leu, CG8827, Networks, MGC130048, determination, ACTG, ACTE, Pflanze, protein, GRO:0005338, BSK, Bsk, Continental Population Groups, csp2, Act5, viridiplantae, L-Leucine, l(1)G0420, Membrane Tissues, Arabidopsis thalianas, halite, ATP hydrolysis-dependent activity, exact), Racial Group, Population Group, protein aggregate, Leucin, foton, bsk, Family Life Cycle, Stock, multicellular organismal biosynthetic process, Group, single organism cellular localization, single-organism biosynthetic process, (Ca2+ + Mg2+)-ATPase, Mouse-ear Cress, membrane region, Inverse Polymerase Chain Reaction, localization within cell, Groups, plants, cloruro sodico, TNFSF14, phototropin 2, PTK4, Racial Stock, act 42A, Reaction, signaling process, Ac5C, CG4027, gamma-sarcoglycan, c-Jun, NON PHOTOTROPIC HYPOCOTYL 1-LIKE, Family Life Cycles, Arabidopses, DmelCG9063, ATP, gamma, br31, Membrane Tissue, NPL1, D-junk, rock salt, l(1)G0330, ATP hydrolase activity, hydrogen ion transport, SG-gamma, BSK/IYK, A. thaliana, membranous organ component, Rek7, LACH1, results, K3G17_4, ATPase-dependent activity, untranslated region, Alpha-1-antichymotrypsin His-Pro-less, DFNA26, Genetic Materials, overlap, Racial Stocks, Filiation, GAT, DFNA20, Genetic Material, region, cellular localisation, Visible Radiations, ATP hydrolysis coupled proton transport, Transphosphorylases, Visible Radiation, membrane of organ, Nested Polymerase Chain Reaction, Spalt, l(1)G0117, Plant, Actin, Race, establishment and maintenance of cellular localization, natrii chloridum, gamma (35kDa dystrophin-associated glycoprotein), beta-actin/Bap47, l(1)G0486, Life Cycles, l(1)G0245, SNF1-RELATED PROTEIN KINASE 2-2, race, Arabidopsis, DMDA, l(1)G0009, ACTL3, Lach1, Cistron, RACE, Protein Kinase, BSK/DJNK, biological signaling, l(1)G0010, Family Member, SGCG_HUMAN, establishment and maintenance of localization in cell or cell membrane, L Isomer, Races, Gene, CG12051, Network, Serpin A3, Inverse, l(2)br31, Ximpact, TYPE, DAGA4, l(1)G0025, Inverse PCR, polypeptide chain, Hek7, integral component of membrane, 35DAG, AW125296, SCG3, Phosphotransferase, Cardaminopsis., study, l34Eb, act5C, complex V (mitochondrial electron transport), Genetic, single-organism cellular localization, Research, formation, cell elongation, chlorure de sodium, ATPase activity, Visible, Transphosphorylase, ATP-protein phosphotransferase, AI854630, Act42a, SNRK2-2, 1700027G07Rik, sels, region of membrane, K21L19_6, site, l(2)34Eb, Mouse ear, Kinship Networks, Family, Cte-II, l(1)G0177, F13M22.21, membrane, T DNA, Jnk, JNK, protein complex, sales, adenosine 5'-triphosphatase activity, Cistrons, impact-a, Phosphotransferases, LGMD2C, jnk, CAD, natural protein, MCK7_2, Protein, whole membrane, sequence, F13M22_21, table salt, Continental, ACTA3, Sal, Radiations, l(1)G0079, Anchored, transmembrane, SPK-2-2, Exhibit, Reactions, Nested, hydrogen transmembrane transport, Leucine, Kinases, INS, VSCM, Photoradiation, act 5C, cyp26, BG:DS08220.3, Salze, Membrane, Population Groups, Mouse-ear Cresses, introduction, LTg, sal, Protein Gene Products, UR-2-R, SCARMD2, Ethnicity, Nested PCR, assay, RWDD5, RAK, Actin5C0PXD039040falsePhototropin2 3'UTR overlaps with the AT5G58150 gene encoding an inactive RLK kinaseThis study aimed to investigate the biological implications of two overlapping sequences in the Arabidopsis genome, the 3UTR of the PHOT2 gene and a putative AT5G58150 gene, encoded on the complementary strand. AT5G58150 is a probably inactive protein kinase that belongs to the transmembrane, leucine rich repeat receptor like kinase family. Phot2 is a UV/blue light, membrane-bound photoreceptor kinase. Thus, both proteins share their cellular localization, apart from the loci proximity. Both the at5g58150 T-DNA SALK_093781C and 35S::AT5G58150-GFP lines overexpress the AT5G58150 gene. However, a detailed analysis did not prove any substantial impact of PHOT2 or AT5G58150 on their mutual expression levels in the different light and osmotic stress conditions. This may result from only a small overlap found for 3UTR regions of both genes consisting of 66 bp. AT5G58150 is a plasma membrane protein, with no apparent kinase activity. We could not observe AT5G58150 homodimer formation or any significant interaction with PHOT2. Our findings show that AT5G58150 may be involved in the development of root structures such as primordia or lateral roots. In line with this, mass spectrometry analysis identified proteins with ATPase activity, which are involved in proton transport and cell elongation, as putative interactors of AT5G58150. Membrane kinases, including other members of the LRR RLK family and BSK kinases (positive regulators of brassinosteroid signalling), can also act as partners for AT5G58150.Wed Dec 21 08:42:00 GMT 2022MSV00009094538238701