Co-sedimentation of Actin, Tubulin andMembranes in the Cytoskeleton Fractions from Peas and Mouse 3T3 Cells

Abe, Shunnosuke; Itō, Yoko; Davies, Eric

doi:10.1093/jxb/43.7.941

Cited by 37 publications

(32 citation statements)

References 23 publications

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“…Frozen seedlings in amount of 0.5 g were ground under liquid nitrogen using cooled mortar and pestle and suspended in about 3 ml of buffer U containing 2 % PTE (polyoxyethylene-10-tridecyl ether, a non-ionic detergent); 1 % DOC (sodium deoxycholate); 200 mM TrisHCl, pH 8.5; 50 mM potassium acetate; 25 mM magnesium acetate; 2 mM EGTA; 100 mg/ml heparin and centrifuged at 12,0009g for 20 min (Abe et al 1992). The obtained supernatants, containing total extractable fraction of polyribosomes, were placed in the centrifuge tubes, on top of 0.5 ml of 50 % ''sucrose cushion''.…”

Section: Polysome Evaluationmentioning

confidence: 99%

The effect of seed conditioning, short-term heat shock and salicylic, jasmonic acid or brasinolide on sunflower (Helianthus annuus L.) chilling resistance and polysome formation

2014

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The aim of this study was to develop the method for increasing resistance of sunflower seedlings 'Wielkopolski' to chilling. Seeds were conditioned at 25°C for 2 days in water to 15, 20 and 25 % moisture content or in salicylic or jasmonic acid in concentration of 10 -2 ; 10 -3 and 10 -4 M or brassinolide in concentration of 10 -6 ; 10 -8 and 10 -10 -15 % moisture content. After 2 days of incubation the conditioned seeds were heat shocked at 45°C for 0, 30, 60, 120 and 240 min and 5 mm seedlings were exposed to chilling at 0°C for 21 days. The effectiveness of the methods was assessed by evaluation of roots growth in Phytotoxkit Microbiotest, changes in the activity of dehydrogenases, the integrity of the cytoplasmic membrane and formation of polysomes after seedling were returned to 25°C for 72 h. Seeds were conditioned at 25°C for 2 days in water to 15 % moisture content and then heat shocked at 45°C for 2 h decreased chilling injury of seedlings expressed by subsequent growth of the roots, electrolyte leakage, dehydrogenases activity and polysomes formation. Application of heat shock of 45°C for 2 h during seed conditioning additionally provided seedling protection against subsequent chilling conditions. Brasinolide, salicylic acid or jasmonic acid applied during seeds conditioning exhibited further beneficial effect on seedling resistance to chilling. The most pronounced effect was obtained due to seed conditioning to 15 % moisture content in solutions of brassinolide in concentration of 10 -8 M. After 2 days of imbibition treated in this way seeds were exposed to heat shock at 45°C for 2 h. The role of physiological events in improvement of sunflower chilling tolerance are discussed.

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Section: Polysome Evaluationmentioning

confidence: 99%

The effect of seed conditioning, short-term heat shock and salicylic, jasmonic acid or brasinolide on sunflower (Helianthus annuus L.) chilling resistance and polysome formation

2014

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“…This buffer allows sequential isolation of four polysome populations [26]: FP, MBP, CBP and CMBP. The supernatant polysomes included FP and, to prevent their degradation by RNAse, the samples were adjusted to buffer U, consisting of 200 mM Tris-HCl, pH 8.5, 50 mM KOAc (potassium acetate), 25 mM Mg(OAc) 2 , 2 mM EGTA, 100 mg/ml heparin, 2% PTE (polyoxyethylene-10-tridecyl ether, a non-ionic detergent) and 1% DOC (sodium deoxycholate) [27] by adding 1/4 vol. of 4× concentrated of buffer U and held on ice prior to further processing.…”

Section: Polysome Isolationmentioning

confidence: 99%

“…All supernatant fractions (FP, MBP, CBP, CMBP) were layered on a 0.5 ml "pad" of 50% (w/v) sucrose in buffer B [50 mM Tris-HCl, pH 7.5, 20 mM KOAc, 10 mM Mg(OAc) 2 ] and centrifuged for 90 min at 300000 × g in Beckman 65 Ti rotor. Sprouts or early seedlings of pea were also homogenized in buffer U [27] to solubilize total polysomes.…”

Section: Polysome Isolationmentioning

confidence: 99%

Effect of osmotic stress on in vitro translational capacity of polysomes and on the composition of polysome-associated proteins in germinating seeds of pea (Pisum sativum L.)

Brosowska-Arendt

Weidner

2012

Acta Soc Bot Pol

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IntroductionChanges in the environment, such as drought, salinity, high or low temperature, are an important factor which affects the growth of crops and the volume of crop yields [1][2][3]. Very important legume grown and consumed extensively worldwide is pea [4]. As a rich source of proteins, carbohydrates, fibre, vitamins and minerals, peas are important in human nutrition [5]. Pea is the fourth leading legume in terms of consumption in the world after soybean, peanuts and bean [6]. One of the major abiotic stress often occurs in Europe is osmotic stresses induced with polyethylene glycol (within −0.5 MPa). The earliest metabolic change caused by water stress is a decreased amount of polysomes [7]. Once the polysomes have been disaggregated, the plant growth is inhibited due to a slower protein synthesis rate [7][8][9]. Considerable reduction in the amount of polysomes in plant tissues is observable after osmotic stress lasting for just 20-30 minutes [10]. Decrease of the content of polysomes in response to abiotic stresses is connected with the process of "switching" the expression of genes from those participating in the growth and development of plants under unstressed conditions to the ones active in response to stress [11].In plant tissues, polysomes can occur as free polysomes (FP), endoplasmic reticulum membrane-bound polysomes (MBP) [12], cytoskeleton-bound polysomes (CBP) [13,14] and cytoskeleton-membrane-bound polysomes (CMBP) [15]. Changes in the distribution of polysomes between the particular fractions reflect changes in the complement of proteins, as each population of polysomes is engaged in the synthesis of specific proteins [16]. Thus, the FP population is mainly involved in the synthesis of soluble proteins of the cytoplasm, cellular nucleus, mitochondria and peroxisomes [17]; the MBP population is engaged in the production of secretory proteins, lysosome proteins and the proteins which are an integral component of the plasmatic membrane and intracellular membranes, including the endoplasmic reticulum [16,17]; finally, the CBP and CMBP populations are responsible for the synthesis of cytoskeleton and stress proteins [16,[18][19][20][21]. The largest share in the total polysome pool consists of the CBP population, which sometimes reaches 70% of the total polysome content [22]. Polysomes contain mRNAs, which may undergo selective translation, result in modification of protein synthesis in response to stress conditions in plants [23,24].The objective of this study was to indicate differences in the composition of polysome-bound proteins and in the products of in vitro translation in pea (Pisum sativum L.) seeds germinating under unstressed conditions and under long-and short-term osmotic stress (−0.5 MPa) followed by post-stress recovery. AbstractPlant growth throughout the world is often limited by unfavourable environmental conditions. This paper reports results of a study on long-and short-term osmotic stress (−0.5 MPa) followed by a recovery on in vitro translational capacity of polysomes a...

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“…This buffer allows sequential isolation of four polysome populations (Davies and Abe 1995): FP, MBP, CBP and CMBP. The supernatant polysomes included the FP and, to prevent their degradation by RNase, the samples were adjusted to buffer U, consisting of 200 mM Tris-HCl, pH 8.5, 50 mM KOAc, 25 mM Mg(OAc) 2 , 2 mM EGTA, 100 mg/ml heparin, 2% PTE (polyoxyethylene-10-tridecyl ether, a non-ionic detergent) and 1% sodium deoxycholate (Abe et al 1992) by adding vol. of 49 concentrated buffer U and held on ice prior to further processing.…”

Section: Polysome Isolation and Quantificationmentioning

confidence: 99%

“…Embryos, sprouts or early seedlings of pea were also homogenized in buffer U (Abe et al 1992) to solubilize total polysomes. The subunits, monosomes and polysomes were purified (using 50% sucrose pad) and the resuspended polysomes were layered on linear 15-60% sucrose gradients and centrifuged in the same way as mentioned above.…”

Section: Polysome Isolation and Quantificationmentioning

confidence: 99%

Effect of osmotic stress on the formation of a population of polysomes and their stability in pea (Pisum sativum L.) seeds

Brosowska-Arendt

Weidner

2011

Acta Physiol Plant

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Plants growing under natural conditions are constantly exposed to various stress factors, which can restrain their productivity and limit yields. This paper deals with the effect of long-and short-term osmotic stress followed by recovery on the formation of polysomes and their stability during germination of pea (Pisum sativum L.) seeds. By isolating polysomes, it is possible to obtain an index which evidences the ability of tissues to synthesize proteins. Changes in the distribution of polysomes often precede measurable changes in amounts of proteins. Under osmotic stress, the dominant population of polysomes was the population of free polysomes (FP). The share of membrane-bound polysomes (MBP) and cytoskeletonbound polysomes (CBP) and cytoskeleton-membranebound polysomes (CMBP) in the total fraction of ribosomes increased under intensive (-1.0 and -1.5 MPa) osmotic stress. These results can suggest that the bound forms of polysomes play an important role in the synthesis of stress proteins. In addition, the stability of polysomes isolated from pea early seedlings growing under unstressed control and osmotic stress conditions was tested. It turned out that polysomes formed under osmotic stress conditions (especially the CMBP) were more resistant to the activity of exogenous ribonucleases than the polysomes in the control samples. Under stress conditions it is highly likely that ribosomes become more densely packed on mRNA thus making it more resistant to ribonuclease. This is just one of the many mechanisms regulating stability of mRNA.

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Co-sedimentation of Actin, Tubulin andMembranes in the Cytoskeleton Fractions from Peas and Mouse 3T3 Cells

Cited by 37 publications

References 23 publications

The effect of seed conditioning, short-term heat shock and salicylic, jasmonic acid or brasinolide on sunflower (Helianthus annuus L.) chilling resistance and polysome formation

The effect of seed conditioning, short-term heat shock and salicylic, jasmonic acid or brasinolide on sunflower (Helianthus annuus L.) chilling resistance and polysome formation

Effect of osmotic stress on in vitro translational capacity of polysomes and on the composition of polysome-associated proteins in germinating seeds of pea (Pisum sativum L.)

Effect of osmotic stress on the formation of a population of polysomes and their stability in pea (Pisum sativum L.) seeds

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