Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2000 Oct 10;97(21):11303-6.
doi: 10.1073/pnas.97.21.11303.

Distinct but overlapping roles of histone acetylase PCAF and of the closely related PCAF-B/GCN5 in mouse embryogenesis

T Yamauchi  1 , J Yamauchi, T Kuwata, T Tamura, T Yamashita, N Bae, H Westphal, K Ozato, Y Nakatani
Affiliations

Distinct but overlapping roles of histone acetylase PCAF and of the closely related PCAF-B/GCN5 in mouse embryogenesis

T Yamauchi et al. Proc Natl Acad Sci U S A. .

Abstract

PCAF plays a role in transcriptional activation, cell-cycle arrest, and cell differentiation in cultured cells. PCAF contributes to transcriptional activation by acetylating chromatin and transcription factors through its intrinsic histone acetylase activity. In this report, we present evidence for the in vivo function of PCAF and the closely related PCAF-B/GCN5. Mice lacking PCAF are developmentally normal without a distinct phenotype. In PCAF null-zygous mice, protein levels of PCAF-B/GCN5 are drastically elevated in lung and liver, where PCAF is abundantly expressed in wild-type mice, suggesting that PCAF-B/GCN5 functionally compensates for PCAF. In contrast, animals lacking PCAF-B/GCN5 die between days 9.5 and 11.5 of gestation. Normally, PCAF-B/GCN5 mRNA is expressed at high levels already by day 8, whereas PCAF mRNA is first detected on day 12.5, which may explain, in part, the distinct knockout phenotypes. These results provide evidence that PCAF and PCAF-B/GCN5 play distinct but functionally overlapping roles in embryogenesis.

PubMed Disclaimer

Figures

Figure 1
Figure 1
_targeted disruption of the PCAF locus. (A) _targeting strategy for PCAF gene. The configuration of the wild-type (WT) allele is shown (Top). The exon encoding the N terminus of PCAF (shown by a box) was replaced by a neomycin resistance gene cassette in the _targeting vector (Middle). The thymidine kinase (tk) gene was used for negative selection. The structure of the _targeted allele is shown (Bottom). The location of the probe used for the Southern blot in B is indicated. Restriction sites: B, BamHI; E, EcoRI; EV, EcoRV; H, HindIII; X, XhoI. kb, kilobase. (B) Southern blot of EcoRV-digested genomic DNAs isolated from offspring of heterozygous mice. The probe hybridizes with 12.5- and 3.5-kb fragments in the wild-type and _targeted allele, respectively. Typical results obtained from PCAF+/+, PCAF+/−, and PCAF−/− mice are shown. (C) Immunoblot of protein extracts from lung of PCAF+/+, PCAF+/−, and PCAF−/− embryos. PCAF could not be detected in PCAF−/− mice (Upper). As an internal control, TFIIB was also detected (Lower).
Figure 2
Figure 2
_targeted disruption of the PCAF-B/GCN5 locus. (A) _targeting strategy for PCAF-B/GCN5 gene. The configuration of the wild-type (WT) allele is shown (Top). The genomic region containing exons encoding the 3′ terminus of PCAF-B/GCN5 (shown by boxes) was replaced by a neomycin resistance gene cassette in the _targeting vector (Middle), yielding the _targeted allele (Bottom) by homologous recombination. Restriction sites: Bg, BglII; E, EcoRI; X, XhoI. (B) Southern blot of EcoRI-digested genomic DNAs isolated from offspring of heterozygous mice. The probe hybridizes with 13- and 10-kb fragments in the wild-type and _targeted allele, respectively. (C) Immunoblot of protein extracts from PCAF-B/GCN5+/+, PCAF-B+/−, and PCAF-B−/− embryos. PCAF-B/GCN5 could not be detected in PCAF-B−/− embryos (Upper). Note that an immunoreactive band around 55 kDa could not be detected. Further characterization would be required to resolve the issue of the N-terminally truncated form of mouse PCAF-B/GCN5. As an internal control, TFIIB was also detected (Lower).
Figure 3
Figure 3
Developmental retardation of PCAF-B/GCN5−/− embryos. Lateral (A) and dorsal (B) views of wild-type (Left) and homozygous (Right) littermates on E9.5. (Bar = 0.5 mm.)
Figure 4
Figure 4
Overproduction of PCAF-B/GCN5 in PCAF null-zygous mice. Immunoblot analyses of protein extracts from brain, lung, and liver of PCAF+/+, PCAF+/−, and PCAF−/− mice were performed. No PCAF was detected in tissues isolated from PCAF−/− mice (Top), whereas PCAF-B/GCN5 was significantly up-regulated in lung and liver of PCAF−/− mice (Middle). TFIIB levels were determined as an internal control (Bottom).
Figure 5
Figure 5
In situ analysis of PCAF and PCAF-B/GCN5 mRNAs in mouse embryos. Dark-field micrographs of parasagittal sections of E10.5 (B–E) and E12.5 (G and H) mouse embryos after hybridization with a PCAF-specific antisense probe (B and E), a PCAF-B/GCN5-specific antisense probe (B and G), or the respective sense probes as a control (C and E). Bright-field micrographs of toluidine blue-stained sections of E10.5 (A) and E12.5 (F) are also shown. Abbreviations: fb, forebrain; h, heart; hb, hindbrain; hl, hind limb; li, liver; lu, lung; nt; neural tube; skm, skeletal muscle. (Bar = 1 mm.)
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Yang X J, Ogryzko V V, Nishikawa J, Howard B H, Nakatani Y. Nature (London) 1996;382:319–324. - PubMed
    1. Kuo M H, Allis C D. BioEssays. 1998;20:615–626. - PubMed
    1. Struhl K. Genes Dev. 1998;12:599–606. - PubMed
    1. Workman J, Kingston R E. Annu Rev Biochem. 1998;67:545–579. - PubMed
    1. Schiltz R L, Nakatani Y. Biochim Biophys Acta. 2000;1470:M37–M53. - PubMed

LinkOut - more resources

  NODES
INTERN 3
Note 1
twitter 2