Product info:

Actin is a key regulator of RNA polymerase (Pol) II-dependent transcription. Positive transcription elongation factor b (P-TEFb), a Cdk9/cyclin T1 heterodimer, has been reported to play a critical role in transcription elongation. However, the relationship between actin and P-TEFb is still not clear. In this study, actin was found to interact with Cdk9, a catalytic subunit of P-TEFb, in elongation complexes. Using immunofluorescence and immunoprecipitation assays, Cdk9 was found to bind to G-actin through the conserved Thr-186 in the T-loop. Overexpression and in vitro kinase assays showed that G-actin promotes P-TEFb-dependent phosphorylation of the Pol II C-terminal domain. An in vitro transcription experiment revealed that the interaction between G-actin and Cdk9 stimulated Pol II transcription elongation. ChIP and immobilized template assays indicated that actin recruited Cdk9 to a transcriptional template in vivo and in vitro. Using cytokine IL-6-inducible p21 gene expression system, we revealed that actin recruited Cdk9 to endogenous gene. Moreover, overexpression of actin and Cdk9 increased histone H3 acetylation and acetylized histone H3 binding to a transcriptional template through the interaction with histone acetyltransferase, p300. Taken together, our results suggested that actin participates in transcription elongation by recruiting Cdk9 for phosphorylation of the Pol II C-terminal domain, and the actin-Cdk9 interaction promotes chromatin remodeling.

Alternative names:

H3.4; H3/g; H3FT; H3t; HIST3H3; Histone H3; HIST1H3A

Species reactivity:

Human, Mouse, Rat, Other (Wide Range)

Host:

Rabbit

Cellular localisation:

Chromosome, Nucleus

Isotype:

IgG

Immunogen:

A synthetic methylated peptide corresponding to residues surrounding K27 of human histone H3

Positive control:

HeLa, NIH/3T3, H3 protein

AA Sequence:

Email info@sobekbio.com for sequence

Purification:

Affinity purification

Molecular weight:

Calculated MW: 15kDa Observed MW: 17KDa

Form:

Liquid

Applications:

IF (Human embryonic stem cells, Mouse E16.5 neocortex, Goat sperm, HEK293T, Rice, Gallus gallus, Homo sapiens, Mus musculus) ChIP (A549, Mouse embryonic stem cells, HeLa, Human, Rice, Gossypium, Oryza sativa, Homo sapiens, Rattus norvegicus) IHC (Mouse lung tumor, Mus musculus, Sus scrofa, Homo sapiens) ChIP-qPCR (Mus musculus, Homo sapiens) IP (Gossypium, Mus musculus) ChIP-Seq (Zea mays) WB (Mus musculus, Homo sapiens, Rattus norvegicus)

Dilution:

WB 1:500 - 1:2000 IHC 1:50 - 1:200 IF 1:50 - 1:200 IP 1:50 - 1:200 ChIP 1:20 - 1:100 ChIP-seq 1:20 - 1:100

Storage buffer:

Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3.

Storage:

Store at 4°C short term. For long-term storage, aliquot and store at -20°C or below. Stable for 12 months at -20°C. Avoid repeated freeze-thaw cycles.

More info:

Email: info@sobekbio.com

Orders:

Email: orders@sobekbio.com