Regulation of an epitope-tagged recombinant Rsk-1 S6 kinase by phorbol ester and erk/MAP kinase

Grove, Jeffrey; Price, Daniel J.; Banerjee, Papia; Balasubramanyam, Ashok; Ahmad, Mir F.; Avruch, Joseph

doi:10.1021/bi00081a018

Cited by 66 publications

(59 citation statements)

References 35 publications

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“…RSK2 was excised from pMT2-HA-RSK2 using XhoI and KpnI and replaced with the PCR product digested with the same enzymes. HA-RSK1 (rat) in pMT2 (46) was kindly provided by Dr. Joseph Avruch. HA-MSK1 in pMT2 is described in Ref.…”

Section: Methodsmentioning

confidence: 99%

Functional Characterization of Human RSK4, a New 90-kDa Ribosomal S6 Kinase, Reveals Constitutive Activation in Most Cell Types

Dümmler

Hauge

Silber

et al. 2005

Journal of Biological Chemistry

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The 90-kDa ribosomal S6 kinases (RSK1-3) are important mediators of growth factor stimulation of cellular proliferation, survival, and differentiation and are activated via coordinated phosphorylation by ERK and 3-phosphoinositide-dependent protein kinase-1 (PDK1). Here we performed the functional characterization of a predicted new human RSK homologue, RSK4. We showed that RSK4 is a predominantly cytosolic protein with very low expression and several characteristics of the RSK family kinases, including the presence of two functional kinase domains and a C-terminal docking site for ERK. Surprisingly, however, in all cell types analyzed, endogenous RSK4 was maximally ( cdc2 -inhibitory kinase Myt1 (7) and the protein kinase Bub1 (8), respectively. In somatic cell types, RSK may stimulate cell division through phosphorylation of substrates like p27 Kip1 (9) and glycogen synthase kinase-3 (10 -12); protein synthesis and cell growth through substrates like elongation factor 2 kinase (13), glycogen synthase kinase-3 (10 -12), transcription initiation factor IA (14), and tuberous sclerosis complex-2 protein (15); cell survival through Bad (16); and transcription through substrates like c-Fos (17) and estrogen receptor (18). Experiments with PC12 cells suggest that RSK is a key mediator of ERK in neurotrophin-induced neuronal differentiation (19). Finally genetic evidence from human and mouse has identified an important role for RSK2 in osteoblast differentiation and function through phosphorylation of activating transcription factor-4 (20) and in stimulation of white adipose tissue mass via an unknown mechanism (21). RSK is related to the mitogen-and stress-activated protein kinases (MSK1 and MSK2), which also contain two kinase domains. MSK, however, is activated by the ERK family as well as by p38 family MAP kinases and thereby functions in signal transduction of growth factors as well as of cellular stress stimuli like UV/radioactive irradiation and proinflammatory cytokines (22,23).The substrates of RSK are phosphorylated by the N-terminal kinase (NTK) domain (24 -27) whose activity is regulated by

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Section: Methodsmentioning

confidence: 99%

Functional Characterization of Human RSK4, a New 90-kDa Ribosomal S6 Kinase, Reveals Constitutive Activation in Most Cell Types

Dümmler

Hauge

Silber

et al. 2005

Journal of Biological Chemistry

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“…The WT mouse akt1, CA mouse akt1 (amino terminus myristoylated), WT rat MEK1, CA rat MEK1 (S218D/S222D), WT rat MEK2, and CA human H-ras (Q61L) cDNA in a pUSEamp vector were purchased from Upstate Biotechnology. The pMT2-HA vector containing rat RSK1 and mouse RSK2 cDNAs (40,41) were kindly provided by Dr. Joe Avruch (Massachusetts General Hospital, Boston, MA) and Dr. Christian Bjorbaek (Beth Israel Deaconess Medical Center, Boston, MA), respectively. The kinase-dead form of RSK1 cDNA (D205N) (42) was generated by mutagenesis.…”

Section: Methodsmentioning

confidence: 99%

Phosphorylation of p27Kip1 at Threonine 198 by p90 Ribosomal Protein S6 Kinases Promotes Its Binding to 14-3-3 and Cytoplasmic Localization

Fujita

Sato

Tsuruo

2003

Journal of Biological Chemistry

179

148

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The cyclin-dependent kinase inhibitor p27Kip1 plays an important role in cell cycle regulation. The cyclin-dependent kinase-inhibitory activity of p27Kip1 is regulated by changes in its concentration and its subcellular localization. Several reports suggest that phosphorylation of p27Kip1 at serine 10, threonine 157, and threonine 187 regulate its localization. We have previously identified that carboxyl-terminal threonine 198 (Thr 198 ) in p27Kip1 is a novel phosphorylation site and that Akt is associated with the phosphorylation at the site (Fujita, N., Sato, S., Katayama, K., and Tsuruo, T. (2002) Kip1 binding to 14-3-3 and its cytoplasmic localization. To prove the direct relationship between 14-3-3 binding and cytoplasmic localization, we constructed a p27Kip1 -R18 fusion protein in which the R18 peptide was fused to the carboxyl-terminal region of p27 Kip1 . The R18 peptide is known to interact with 14-3-3 independent of phosphorylation. The p27 Kip1 -R18 distributed mainly in the cytosol, whereas mutant p27 Kip1 -R18 (p27 Kip1 -R18-K2) that had no 14-3-3 binding capability existed mainly in the nucleus. These results indicate that RSKs play a crucial role in cell cycle progression through translocation of p27 Kip1 , in addition to Akt, to the cytoplasm in a phosphorylationand 14-3-3 binding-dependent manner.

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“…To generate MIAL tagged with the 9-amino-acid hemagglutinin (HA) epitope, cDNA of human MIAL was amplified by PCR using a sense primer complementary to nucleotides Ϫ68 to Ϫ47 from the initiation codon and introducing a NotI site (5Ј-CAGTCGATAGCGGCCGCAAGGCAGGAACCACTGA-AGTC-3Ј) and an antisense primer deleting the natural stop codon and introducing a NotI site (5Ј-GACACGTAGGCGGCCGCCTCGCA-GAAGAAGTCAATATC-3Ј). The PCR product was digested with NotI and inserted in-frame with a C-terminally located HA tag in the mammalian expression vector pMT2 (Grove et al, 1993) using the NotI site. Before subcloning, the pMT2 vector had been modified by excision of the RSK1 gene present in the vector and introduction of a stop codon after the HA tag.…”

Section: Mial Expression Vectorsmentioning

confidence: 99%

Identification and Characterization of an Inner Ear-Expressed Human Melanoma Inhibitory Activity (MIA)-like Gene (MIAL) with a Frequent Polymorphism That Abolishes Translation

et al. 2001

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To discover new cochlea-specific genes as candidate genes for nonsyndromic hearing impairment, we searched in The Institute of Genome Research database for expressed sequence tags isolated from the cochlea only. This led to the cloning and characterization of a human gene named melanoma inhibitory activity-like (MIAL; HGMW-approved symbol OTOR alias MIAL) gene. In situ hybridization revealed MIAL expression in a cell layer beneath the sensory epithelium of cochlea and vestibule of human fetal inner ear. No other human tissue, except fetal brain, showed expression of MIAL when analyzed by in situ hybridization or reverse transcription-polymerase chain reaction. The cDNA of the mouse homologue was also cloned and mapped about 80 cM from the top of mouse chromosome 2. In mouse, Mial was also expressed in the cochlea and the vestibule of the inner ear, as well as in brain, eye, limb, and ovary. Expression in mammalian cell cultures showed that MIAL is translated as an ϳ15-kDa polypeptide that is assembled into a covalently linked homodimer, modified by sulfation, and secreted from the cells via the Golgi apparatus. In the human MIAL gene, a frequent polymorphism was discovered in the translation initiation codon (ACG instead of ATG). Of 505 individuals, 48 (9.5%) were ATG/ ACG heterozygous and 1 (0.2%) was homozygous for ACG. No MIAL protein was synthesized in cells transfected with cDNA of the ACG allele. The inner earrestricted expression pattern and the existence of an inactive allele suggest that MIAL may contribute to inner-ear dysfunction in humans.

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Regulation of an epitope-tagged recombinant Rsk-1 S6 kinase by phorbol ester and erk/MAP kinase

Cited by 66 publications

References 35 publications

Functional Characterization of Human RSK4, a New 90-kDa Ribosomal S6 Kinase, Reveals Constitutive Activation in Most Cell Types

Functional Characterization of Human RSK4, a New 90-kDa Ribosomal S6 Kinase, Reveals Constitutive Activation in Most Cell Types

Phosphorylation of p27Kip1 at Threonine 198 by p90 Ribosomal Protein S6 Kinases Promotes Its Binding to 14-3-3 and Cytoplasmic Localization

Identification and Characterization of an Inner Ear-Expressed Human Melanoma Inhibitory Activity (MIA)-like Gene (MIAL) with a Frequent Polymorphism That Abolishes Translation

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