Heat shock modulates phosphorylation status and activity of nucleoside diphosphate kinase in cultured sugarcane cells.
Nucleoside diphosphate kinase (NDPK) is involved in the regeneration of nucleoside triphosphates (NTPs) through its phosphotransferase activity via an autophosphorylating histidine residue. Additionally, autophosphorylation of serine and/or threonine residues is documented for NDPKs from various organisms. However, the metabolic significance of serine/threonine phosphorylation has not been well characterized. In this study we report the cloning and characterization of NDPKI from cultured sugarcane (Saccharum officinarum L. line H50-7209) cells, and modulation of serine autophosphorylation of NDPK1 in response to heat-shock (HS). Heat-shock treatment at 40°C for 2 h resulted in a 40% reduction in labeled phosphoserine in NDPK1. This dephosphorylation was accompanied by an increase in NDPK enzyme activity. In contrast, NDPK1 in cultured tobacco (cv. W-38) cells did not show changes in autophosphorylation or increased enzyme activity in response to HS. The mRNA or protein level of NDPK1 did not increase in response to HS. Sugarcane cells sustain the constitutive protein synthesis in addition to heat-shock protein synthesis during HS, while constitutive protein synthesis is significantly reduced in tobacco cells during HS. Thus, HS modulation of NDPK1 activity and serine dephosphorylation in sugarcane cells may represent an important physiological role in maintaining cellular metabolic functions during heat stress.
Dharmasiri
Sunethra
S
Department of Biology, Texas State University, 601, University Drive, San Marcos, USA. sd20@txstate.edu
Harrington
H Michael
HM
Dharmasiri
Nihal
N
D000595:Amino Acid Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018869:Heat-Shock Response; D008969:Molecular Sequence Data; D009701:Nucleoside-Diphosphate Kinase / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D031786:Saccharum / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment; D012694:Serine / Q000378:metabolism; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics
D018076:DNA, Complementary; D010940:Plant Proteins; D012694:Serine; D009701:Nucleoside-Diphosphate Kinase
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