Abstract
Thirty-one cDNAs corresponding to pdi genes [inorganic phosphate (Pi) deficiency-inducible genes] were previously isolated through the differential screening of a cDNA library constructed from the mycelium of Pholiota nameko. Among the cDNAs, pdi370 was analyzed here. The deduced amino acid sequence showed high similarity to fungal ribonucleases (RNases) and contained two signature sequences conserved in T2 family RNases: CAS1 and CAS2. Genomic DNA harboring the pdi370 gene was isolated from a genomic library of P. nameko. Sequence analysis showed that the pdi370 gene is interrupted by 16 introns and that the promoter region contains two cis-acting sequences found in Pi deficiency-induced genes from Saccharomyces cerevisiae, together with several known functional elements, such as a TATA box. RNase activity in the mycelium and culture filtrate increased 5.6-fold and 5.2-fold, respectively, under Pi-deficient conditions. Staining for RNase activity showed that at least four RNases are induced and secreted under the conditions. The N-terminal sequence of one of them agreed with that of the pdi370 gene product.
Similar content being viewed by others
References
Abel S, Glund K (1986) Localization of RNA-degrading enzyme activity within vacuoles of cultured tomato cells. Physiol Plant 66:79–86
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
Baldwin JC, Karthikeyan AS, Raghothama KG (2001) LEPS2, a phosphorus starvation-induced novel acid phosphatase from tomato. Plant Physiol 125:728–737
Bariola PA, Howard CJ, Taylor CB, Verburg MT, Jaglan VD, Green PJ (1994) The Arabidopsis ribonuclease gene RNS1 is tightly controlled in response to phosphate limitation. Plant J 6:673–685
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254
Drescher DG, Lee KS (1978) Extraction of fixed, stained protein bands from gels for micro amino acids analysis using o-phthaldialdehyde. Anal Biochem 84:559–569
Goldstein AH, Baertlein DA, Danon A (1989) Phosphate starvation stress as an experimental system for molecular analysis. Plant Mol Biol Rep 7:7–16
Green PJ (1994) The ribonucleases of higher plants. Annu Rev Plant Physiol Plant Mol Biol 45:421–445
Gurr SJ, Unkles SE, Kinghorn JR (1987) The structure and organization of nuclear genes of filamentous fungi. In: Kinghorn JR (ed) Gene structure in eukaryotic microbes. IRL Press, Oxford, pp 93–139
Hahnen E, Znamenskaya L, Koczan D, Leshchinskaya I, Hobom G (2000) A novel secreted ribonuclease from Bacillus intermedius: gene structure and regulatory control. Mol Gen Genet 263:571–580
Hofmann K, Bucher P, Falquet L, Bairoch A (1999) The PROSITE database, its status in 1999. Nucleic Acids Res 27:215–219
Horiuchi H, Yanai K, Takagi M, Yano K, Wakabayashi E, Sanda A, Mine S, Ohgi K, Irie M (1988) Primary structure of a base non-specific ribonuclease from Rhizopus niveus. J Biochem (Tokyo) 103:408–418
Igic B, Kohn JR (2001) Evolutionary relationships among self-incompatibility RNases. Proc Natl Acad Sci USA 98:13167–13171
Inokuchi N, Kobayashi H, Hara J, Itagaki T, Koyama T, Iwama M, Ohgi K, Irie M (2000) Amino acid sequence of a unique ribonuclease with a C-terminus rich in O-glycosylated serine and threonine from culture medium of Lentinus edodes. Biosci Biotechnol Biochem 64:44–51
Irie M, Ohgi K (2001) Ribonuclease T2. Methods Enzymol 341:42–55
Joh T, Tasaki Y, Hara T, Hayakawa T (2001) Induction and secretion of RNA-degrading enzymes by phosphate deficiency in Pholiota nameko. Biosci Biotechnol Biochem 65:2561–2564
Kai M, Masuda Y, Kikuchi Y, Osaki M, Tadano T (1997) Isolation and characterization of a cDNA from Catharanthus roseus which is highly homologous with phosphate transporter. Soil Sci Plant Nutr 43:227–235
Köck M, Löffler A, Abel S, Glund K (1995) cDNA structure and regulatory properties of a family of starvation-induced ribonucleases from tomato. Plant Mol Biol 27:477–485
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Liu C, Muchhal US, Raghothama KG (1997) Differential expression of TPSI1, a phosphate starvation-induced gene in tomato. Plant Mol Biol 33:867–874
Liu C, Muchhal US, Uthappa M, Kononowicz AK, Raghothama KG (1998) Tomato phosphate transporter genes are differentially regulated in plant tissues by phosphorus. Plant Physiol 116:91–99
Löffler A, Abel S, Jost W, Beintema JJ, Glund K (1992) Phosphate-regulated induction of intracellular ribonucleases in cultured tomato (Lycopersicon esculentum) cells. Plant Physiol 98:1472–1478
Ma R-C, Oliveira MM (2000) The RNase PD2 gene of almond (Prunus dulcis) represents an evolutionarily distinct class of S-like RNase genes. Mol Gen Genet 263:925–933
Mukatira UT, Liu C, Varadarajan DK, Raghothama KG (2001) Negative regulation of phosphate starvation-induced genes. Plant Physiol 127:1854–1862
Nielsen H, Engelbrecht J, Brunak S, Heijne G von (1997) Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10:1–6
Nürnberger T, Abel S, Jost W, Glund K (1990) Induction of an extracellular ribonuclease in cultured tomato cells upon phosphate starvation. Plant Physiol 92:970–976
Oshima Y (1997) The phosphatase system in Saccharomyces cerevisiae. Genes Genet Syst 72:323–334
Ozeki K, Kitamoto K, Gomi K, Kumagai C, Tamura G, Hara S (1991) Cloning and nucleotide sequence of the genomic ribonuclease T2 gene (rntB) from Aspergillus oryzae. Curr Genet 19:367–373
Richardson AE (1994) Soil microorganisms and phosphorus availability. In: Pankhurst CE, Doube BM, Gupta VVSR, Grace PR (eds) Soil biota management in sustainable farming systems. CSIRO, Canberra, pp 50–62
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
Shirzadegan M, Christie P, Seemann JR (1991) An efficient method for isolation of RNA from tissue cultured plant cells. Nucleic Acids Res 19:6055
Tasaki Y, A Azwan, Hara T, Joh T (2001) Effect of phosphate deficiency on growth and protein profile in three strains of Pholiota nameko. Mycoscience 42:489–498
Tasaki Y, Kamiya Y, A Azwan, Hara T, Joh T (2002) Gene expression during Pi deficiency in Pholiota nameko: accumulation of mRNAs for two transporters. Biosci Biotechnol Biochem 66:790–800
Wagner DB, Furnier GR, Saghai-Maroof MA, Williams SM, Dancik BP, Allard RW (1987) Chloroplast DNA polymorphisms in lodgepole and jack pines and their hybrids. Proc Natl Acad Sci USA 84:2097–2100
Wasaki J, Omura M, Osaki M, Ito H, Matsui H, Shinano T, Tadano T (1999) Structure of a cDNA for an acid phosphatase from phosphate-deficient lupin (Lupinus albus L.) roots. Soil Sci Plant Nutr 45:439–449
Watanabe H, Fauzi H, Iwama M, Onda T, Ohgi K, Irie M (1995) Base non-specific acid ribonuclease from Irpex lacteus, primary structure and phylogenetic relationships in RNase T2 family enzyme. Biosci Biotechnol Biochem 59:2097–2103
White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, San Diego, pp 315–322
Wilson CM (1975) Plant nucleases. Annu Rev Plant Physiol 26:187–208
Wykoff DD, Grossman AR, Weeks DP, Usuda H, Shimogawara K (1999) Psr1, a nuclear localized protein that regulates phosphorus metabolism in Chlamydomonas. Proc Natl Acad Sci USA 96:15336–15341
Znamenskaya LV, Vershinina OA, Vershinina VI, Leshchinskaya IB, Hartley RW (1999) Expression of the genes for guanyl-specific ribonucleases from Bacillus intermedius and Bacillus pumilus is regulated by the two component signal transduction system PhoP–PhoR in B. subtilis. FEMS Microbiol Lett 173:217–222
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by J. Heitman
Rights and permissions
About this article
Cite this article
Tasaki, Y., Azwan, A., Hara, T. et al. Structure and expression of a phosphate deficiency-inducible ribonuclease gene in Pholiota nameko . Curr Genet 45, 28–36 (2004). https://doi.org/10.1007/s00294-003-0463-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00294-003-0463-7