Identification of the active region of the DNA synthesis inhibitory gene p21Sdi1/CIP1/WAF1.

Nakanishi, Makoto; Robetorye, Ryan S.; Adami, Guy R.; Pereira‐Smith, Olivia M.; Smith, James R.

doi:10.1002/j.1460-2075.1995.tb07031.x

Cited by 172 publications

(125 citation statements)

References 33 publications

(25 reference statements)

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“…p21 can also associate with PCNA (Flores-Rozas et al, 1994) and inhibit DNA synthesis (Noda et al, 1994;Waga et al, 1994) but not DNA repair in vitro Shivji et al, 1994). The PCNAbinding region (around amino acids 142 ± 164) lies at the very C-terminus of p21 (Chen et al, 1995;Goubin and Ducommun, 1995;Luo et al, 1995;Nakanishi et al, 1995;Warbrick et al, 1995), as revealed by the crystal structure of a 22 residue peptide derived from the C-terminus of p21 in complex with PCNA (Gulbis et al, 1996). The PCNA-binding site overlaps the nuclear localization signal (residues 140 ± 156) and the C-terminal cyclin-binding site.…”

Section: Introductionmentioning

confidence: 99%

“…Residues *10 ± 80 of p21, p27 Kip1 and p57 Kip2 are related in sequence, and this region has been identi®ed as the cyclin and CDK-binding region (Chen et al, 1995;Goubin and Ducommun, 1995;Luo et al, 1995;Nakanishi et al, 1995). Within this region, there are separate domains for binding to cyclin (residues 16 ± 24) (Adams et al, 1996;Ball et al, 1997;Chen et al, 1996;Zhu et al, 1995) and to CDK (residues 45 ± 65) (Ball et al, 1997;Goubin and Ducommun, 1995;Nakanishi et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

“…Within this region, there are separate domains for binding to cyclin (residues 16 ± 24) (Adams et al, 1996;Ball et al, 1997;Chen et al, 1996;Zhu et al, 1995) and to CDK (residues 45 ± 65) (Ball et al, 1997;Goubin and Ducommun, 1995;Nakanishi et al, 1995). A second cyclin-binding site on p21 has recently been identi®ed near the C terminus, which includes residues 152 ± 158 (Adams et al, 1996;Ball et al, 1997;Chen et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

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Expression of a novel form of p21Cip1/Waf1 in UV-irradiated and transformed cells

Poon

Hunter

1998

Oncogene

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The tumor suppressor p53 and its target the CDK inhibitor p21 (Cip1/Waf1) are key components of the cellular response to DNA damage. Insight into how p21 is regulated in normal cells, and how it may be deregulated in tumor cells is important for the understanding of tumorigenesis. p21 was induced in normal human diploid ®broblasts after UV irradiationinduced DNA damage, but, at a high dose of UV irradiation, a faster mobility form of p21 on SDS ± PAGE (designated p21D) was expressed. Surprisingly, in a variety of growing transformed cell lines, the level of p21 was low but p21D was prominent. We found that p21D appeared to be derived through a loss of around 10 amino acids from the C-terminus of p21, which theoretically would remove the PCNA binding domain, a second cyclin binding domain and the nuclear localization signal sequence. Several characteristics distinguish p21 from p21D. Both the full length p21 and p21D could be stabilized by a proteasome inhibitor, but only the full length p21 was associated with Cdk2 and PCNA. Consistent with this, gel ®ltration chromatography revealed that all the full length p21 in the cell was complexed to other proteins, whereas a signi®cant portion of p21D was in monomeric form. Moreover, p21 was mainly localized to the nucleus, but p21D was mainly localized to the cytoplasm. We propose that the decrease in p21 and increase in p21D could contribute to the deregulation of the cell cycle, and could be a mechanism involved in cellular transformation.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression of a novel form of p21Cip1/Waf1 in UV-irradiated and transformed cells

Poon

Hunter

1998

Oncogene

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“…The amino end of p21 WAF1/CIP1 contains an inhibitory domain of cdk, which inhibits cyclin D and E associated cdks activities. The carboxyl end of p21 WAF1/CIP1 mediates the PCNA-binding and activity of DNA polymerase d (Chen et al, 1995;Luo et al, 1995;Nakanishi et al, 1995;Warbrick et al, 1995). Our results showed that p21 WAF1/CIP1 inhibited [ 3 H]thymidine incorporation and reduced cdk2 kinase activity in human liver cancer cells but did not affect expression of cdk2 and cdk4.…”

Section: Discussionmentioning

confidence: 67%

A cyclin-dependent kinase inhibitor (p21WAF1/CIP1) affects thymidine incorporation in human liver cancer cells

et al. 2002

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p21 WAF1/CIP1 is a universal cyclin-dependent kinase inhibitor. To investigate the role of p21 WAF1/CIP1 in proliferation of human liver cancer cells, we examined the expression of p53, p21 WAF1/CIP1 , cdk2 and cdk4 expression in two human liver cancer cell lines (HepG2 and PLC/PRF/5 cells). The effects of p21 WAF1/CIP1 on [ 3 H]thymidine incorporation and cdks were also examined in these cells. HepG2 cells expressed all these proteins with lower level of cdk4. PLC/PRF/5 cells expressed the other proteins except p21 WAF1/CIP1 . Transfection of p21 WAF1/CIP1 gene inhibited [ 3 H]thymidine incorporation of both cells with different extent. Although the transfection of p21 WAF1/CIP1 did not affect cdk2 and cdk4 expression, it did reduce cdk2 kinase activity by 20%. These results suggest that: (a) p21 WAF1/CIP1 involved in DNA synthesis of human liver cancer cells; (b) p21 WAF1/CIP1 could be a target gene for the treatment of human hepatocellular carcinoma.

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“…Deletion anal-ysis has identified two separate domains within the 164-amino acid human p21 coding sequence. Residues 1-71 alone are highly active in CDK binding and inhibition, and residues 141-160 efficiently bind PCNA and inhibit SV40 DNA replication in vitro (Chen et al, 1995;Luo et al, 1995;Nakanish et al, 1995). To provide a structural basis for the biochemical properties of p2 I , we have purified recombinant p2 1 on a large scale to near homogeneity.…”

mentioning

confidence: 99%

Purification and crystallization of cyclin‐dependent kinase inhibitor p21

R.¹,

Nichols²,

Ke³

1996

Protein Science

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p21, a universal inhibitor of mammalian cyclin-dependent kinases (CDK), regulates cell cycle progression by forming various distinct protein complexes with cyclins, CDKs, and the proliferating cell nuclear antigen. We have overexpressed recombinant human p21 in E. coli and purified active p2 1 to near homogeneity on a large scale. Crystals of recombinant p21 have been grown in the space group P2, a = 157.4, b = 152.7, c = 90.6 A, and fl = 92.7'. The diffraction data of the recombinant p21 have been collected to 2.5 and 3.5 8, resolution for the native crystal and two heavy atom derivatives of mercury and iridium.Keywords: cell cycle; crystallization; cyclin dependent kinase inhibitor; X-ray diffractionThe primary control of eukaryotic cell cycle progression is provided by the sequential formation, activation, and subsequent inactivation of a family of serinekhreonine protein kinases, consisting of a catalytic subunit, a CDK (cyclin-dependent kinase), and an activating regulatory subunit, a cyclin. The enzymatic activity of a CDK is regulated by at least three different mechanisms: cyclin binding and activation, subunit phosphorylation, and association with and inhibition by a CDK inhibitor (reviewed by Sherr and Roberts, 1995). In mammalian cells, there exist at least two distinct families of CDK inhibitors, represented by two prototype CDK inhibitors, p21 and p16. The p16 family currently includes four isolated genes (reviewed by Sherr and Roberts, 1995): p 1 61NK4" (also variously known as MTSI, CDK41, and CDKN2), pl5INK4' (also known as MTS2, PI^'^''^'), and ~1 9 '~~~~ (reviewed by Sherr and Roberts, 1995). p16 family inhibitors regulate two closely related CDKs (CDK4 and CDK6) through two distinct mechanisms: 1) formation of binary complexes with the catalytic CDK4 or CDK6 to prevent activation by cyclin D binding and

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Identification of the active region of the DNA synthesis inhibitory gene p21Sdi1/CIP1/WAF1.

Cited by 172 publications

References 33 publications

Expression of a novel form of p21Cip1/Waf1 in UV-irradiated and transformed cells

Expression of a novel form of p21Cip1/Waf1 in UV-irradiated and transformed cells

A cyclin-dependent kinase inhibitor (p21WAF1/CIP1) affects thymidine incorporation in human liver cancer cells

Purification and crystallization of cyclin‐dependent kinase inhibitor p21

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