Purification and Activation of Recombinant p38 Isoforms α, β, γ, and δ

Keesler, George A.; Bray, Jeff; Hunt, John B.; Johnson, David A.; Gleason, Tom; Yao, Zhengbin; Wang, Shen-Wu; Parker, Carol; Yamane, Harvey; Cole, Craig N.; Lichenstein, Henri S.

doi:10.1006/prep.1998.0947

Cited by 64 publications

(62 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our results suggest that the more important isoform in the regulation of myocyte hypertrophy is p38β, since only p38β isoform significantly activated gene expression of natriuretic peptides ANP and BNP. These results agree with those of Wang et al, who observed hypertrophic response by p38 overexpression in cardiomyocytes (Wang et al, 1998). In addition, we found that activation of p38β pathway also induced an increase in ANP mRNA levels in vivo.…”

Section: Discussionsupporting

confidence: 94%

“…Adenoviral vectors encoding p38 MAPK pathway proteins were generated as described (Wang et al, 1998) and BNP promoter constructs as reported earlier . Further details about recombinant adenoviruses and BNP constructs are presented in Supplementary Materials and Methods.…”

Section: Adenoviral Vectors and Plasmidsmentioning

confidence: 99%

“…In addition, both upstream MKKs (MKK3 or MKK6) similarly enhanced the activity of the p38 isoforms. However, because prior data suggests that p38α is regulated by MKK3, and p38β by both MKK3 and MKK6 Keesler et al, 1998), we selected to use combinations of WT p38α + MKK3b(E) and WT p38β + MKK6b(E) to maximally induce the activation of p38 isoforms.…”

Section: Function Of Recombinant Adenoviruses and P38 Inhibitor Sb203580mentioning

confidence: 99%

“…Wang et al have suggested that p38α (with MKK3b) might have pro-apoptotic effects while the overexpression of the p38β (with MKK6b) isoform results in a hypertrophic response of cultured cardiac myocytes (Wang et al, 1998). These findings were reiterated by Saurin et al, who demonstrated that inhibition of the p38α isoform, and not p38β, led to an increase in cell viability and protection (Saurin et al, 2000).…”

Section: Introductionmentioning

confidence: 98%

“…Of the four p38 isoforms (α, β, γ and δ), only p38α and p38β isoforms are substantially expressed in heart Li et al, 1996;Jiang et al, 1997). Selective activation of different p38 isoforms by distinct MKKs has also been observed; MKK6, which is 80 % homologous to the isoform MKK3, can activate all four p38 isoforms, whereas MKK3 preferentially activates only p38α, p38γ, and p38δ Keesler et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Distinct regulation of B-type natriuretic peptide transcription by p38 MAPK isoforms

Koivisto

Kaikkonen

Tokola

et al. 2011

Molecular and Cellular Endocrinology

View full text Add to dashboard Cite

Persistent controversy underlies the functional roles of specific p38 MAPK isoforms in cardiac biology and regulation of hypertrophy-associated genes. Here we show that adenoviral gene transfer of p38β but not p38α increased B-type natriuretic peptide (BNP) mRNA levels in vitro as well as atrial natriuretic peptide mRNA levels both in vitro and in vivo. Overexpression of p38α, in turn, augmented the expression fibrosis-related genes connective tissue growth factor, basic fibroblast growth factor and matrix metalloproteinase-9 both in vitro and in vivo. p38β-induced BNP transcription was diminished by mutation of GATA-4 binding site, whereas overexpression of MKK6b, an upstream regulator of p38α and p38β, activated BNP transcription through both GATA-4 and AP-1. Overexpression of MKK3, upstream regulator of p38α, induced BNP transcription independently from AP-1 and GATA-4. These data provide new evidence for diversity in downstream targets and functional roles of p38 pathway kinases in regulation of hypertrophy-associated cardiac genes.

show abstract

Section: Discussionsupporting

confidence: 94%

Section: Adenoviral Vectors and Plasmidsmentioning

confidence: 99%

Section: Function Of Recombinant Adenoviruses and P38 Inhibitor Sb203580mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Distinct regulation of B-type natriuretic peptide transcription by p38 MAPK isoforms

Koivisto

Kaikkonen

Tokola

et al. 2011

Molecular and Cellular Endocrinology

View full text Add to dashboard Cite

show abstract

Differential tissue expression and activation of p38 MAPK α, β, γ, and δ isoforms in rheumatoid arthritis

Korb¹,

Tohidast‐Akrad²,

Cetin³

et al. 2006

Arthritis & Rheumatism

130

View full text Add to dashboard Cite

Objective. Activation of p38 MAPK is a key signaling step in chronic inflammation. Inhibition of p38 MAPK is considered to be a promising future strategy to control inflammatory diseases, but studies of compounds to inhibit this kinase have so far been limited to investigation of their side effects. We undertook the present study to investigate which specific molecule, among 4 different isoforms of p38 MAPK (␣, ␤, ␥, and ␦), is predominantly expressed and activated in inflammation. Such knowledge could allow more specific targeting of p38 MAPK in inflammatory disease.Methods. Studies were performed on inflamed tissue from patients with rheumatoid arthritis, as a prototype of inflammatory disease. The expression and activation of the ␣, ␤, ␥, and ␦ isoforms of p38 MAPK were examined by immunoblotting, immunoprecipitation, and immunohistochemistry.Results. Immunoblot analysis revealed that ␣ and ␥ were the predominantly expressed p38 MAPK isoforms, whereas the other 2 isoforms were less frequently present. By immunohistochemistry, the expression of all p38 MAPK isoforms was localized to the synovial lining layer as well as to blood vessels. Colabeling with cellspecific markers revealed that macrophages expressed the ␣ and ␥ isoforms, synovial fibroblasts the ␤ and ␥ isoforms, and granulocytes the ␦ isoform, whereas T lymphocytes were rarely positive for any p38 MAPK isoform. Double-labeling with isoform-specific antibody and pan-p38 antibody against the phosphorylated form of p38 MAPK showed activation of the ␣ and ␥ isoforms. Occasional activation of the ␤ isoform was also noted in the synovial lining and the endothelium, whereas the ␦ isoform, although expressed in pericytes around blood vessels, was not phosphorylated. This phosphorylation pattern was confirmed in immunoprecipitation studies in which activated p38 MAPK from synovial tissue extracts was identified as p38 MAPK␣ and -␥ but not p38 MAPK␤ or -␦.Conclusion. These data show that the ␣ and ␥ isoforms of p38 MAPK dominate in chronic inflammation. Effective strategies to inhibit p38 MAPK should therefore aim to specifically target either or both of these isoforms.The p38 MAPKs are members of a larger group of serine/threonine protein kinases, which allow the transduction of extracellular stress signals to the cell nucleus. MAPKs are reporters of changes in the extracellular milieu, which lead to cellular responses allowing adaptation to changed physiologic and pathologic circumstances. P38 MAPK is considered to be a common pathway of inflammatory stimuli such as tumor necrosis factor (TNF) and interleukin-1 (IL-1), as well as environmental stresses such as heat stress, osmotic shock, ultraviolet light, and cytotoxic chemicals (1,2). Thus, p38 MAPKs function as an "emergency switch" that allows a broad cellular response by turning on the target genes of transcription factors, cytokines, and their surface receptors. These proteins are therefore considered to be a

show abstract

Modulation of human cytomegalovirus immediate‐early gene enhancer by mitogen‐activated protein kinase kinase kinase‐1

Sun¹,

Harrowe²,

Reinhard³

et al. 2001

J of Cellular Biochemistry

View full text Add to dashboard Cite

The immediate-early (IE) promoter of human cytomegalovirus (HCMV) constitutes a primary genetic switch, which determines the progression of viral infection. Earlier reports by others have shown mitogen-activated protein kinase kinase kinase-1 (MEKK1) to be able to up-regulate HCMV-IE promoter through downstream mitogen-activated protein kinase (MAPK) pathways. However, we noticed that the activation of the HCMV-IE promoter by constitutively active MEKK1 (MEKK1-TRU) might not be through the MAPK pathways. Using a HCMV-IE enhancer/promoter (- 522 to + 72) driving a luciferase reporter, we demonstrated that the downstream MAPK activation actually repressed the up-regulation of the promoter by MEKK1 in CHO-K1 and human 293 cells. We further found that the up-regulation of HCMV-IE promoter by MEKK1 could be in great extent suppressed by over-expression of IkappaBalpha. Deletion of the NFkappaB/rel sites in the HCMV-IE enhancer region by mutagenesis proportionally reduced the transcriptional activation by MEKK1-TRU, whereas deletion of the ATF/CREB binding sites or cyclic AMP response elements (CRE) had no effects. Furthermore, the NFkappaB/rel deletion mutant also showed repression on the basic transcription activity of the HCMV-IE promoter. Our results indicate that the NFkappaB/rel sites are not only responsible for the modulation of HCMV-IE enhancer activity by MEKK1 but also control the basic transcription activity of the HCMV-IE promoter. On the other hand, the four consensus CRE sites were found to have no function in the activation of the promoter by MEKK1.

show abstract

Purification and Activation of Recombinant p38 Isoforms α, β, γ, and δ

Cited by 64 publications

References 25 publications

Distinct regulation of B-type natriuretic peptide transcription by p38 MAPK isoforms

Distinct regulation of B-type natriuretic peptide transcription by p38 MAPK isoforms

Differential tissue expression and activation of p38 MAPK α, β, γ, and δ isoforms in rheumatoid arthritis

Modulation of human cytomegalovirus immediate‐early gene enhancer by mitogen‐activated protein kinase kinase kinase‐1

Contact Info

Product

Resources

About