Phosphorylation of human growth hormone by the epidermal growth factor-stimulated tyrosine kinase.

Baldwin, Graham S.; Grego, Boris; Hearn, Milton Thomas William; Knesel, Jaromir; Morgan, Francis J.; Simpson, Richard J.

doi:10.1073/pnas.80.17.5276

Cited by 27 publications

(5 citation statements)

References 32 publications

(44 reference statements)

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“…Near the end of this cluster are 5 consecutive glutamic acid residues followed by a tyrosine. This sequence is very similar to the v-src tyrosine phosphorylation site found in several proteins including polyoma middle T antigen (22). Recently, it has been reported that several topoisomerases can be phosphorylated by tyrosine protein kinases with concomitant loss of activity (23).…”

Section: Agaagatgaagccaccattcacaagagaattattgatagagaaattgaaaaatatcagcgmentioning

confidence: 62%

Cloning, characterization, and sequence of the yeast DNA topoisomerase I gene.

Thrash¹,

Bankier²,

Barrell³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

174

105

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The structural gene for yeast DNA topoisomerase I (TOP)) has been cloned from two yeast genomic plasmid banks. Integration of a plasmid carrying the gene into the chromosome and subsequent genetic mapping shows that TOPI is identical to the gene previously called MAKI. Seven top) (maki) mutants including gene disruptions are viable, demonstrating that DNA topoisomerase I is not essential for viability in yeast. A 3787-base-pair DNA fragment including the gene has been sequenced. The protein predicted from the DNA sequence has 769 amino acids and a molecular weight of90,020. MATERIALS AND METHODSEscherichia coli Strains and Media. E. coli K-12 strain JA221 trpES leuB6 lacY recA thi hsdR was used for plastnid propagation. It was grown in L broth (4) with 50 ,jg of ampicillin per ml. Strain JA194 trpC leuB grown on M9 medium (4) supplemented with 5 ,g of tryptophan per ml and 50 pug of ampicillin per ml was used to screen for LEU2 plasmids; the yeast LEU2 gene complements the E. coli leuB mutation. Plasmids were prepared by an alkaline lysis protocol (4).Yeast Strains and Media. The following S. cerevisiae strains were used: CT552: MATa ura 3-52 his3 his7 leu2-3,112 trpl ade2-101 can]; SD2a: MATa ade2 ura3 leu2 his3 trpl canl topl-I (makl-l) top2-1; SD2c: MATa ade2 ura3 his3 his7 leu2 can] topl-l; CT553: MATa leu2 lysll his] ura3 trpl pet] 7; the diploid CT554 MATa/a leu2/leu2 trpl/+ lys]l /+ pet]7/+ hisl/+ adel/+ ura3/+. All the above strains were constructed in this laboratory using strains described previously (1, 2). Other yeast strains used include 6287-20B: MATa karl-i leu2 from F. Winston (5); 418D:cir0 MATa leu2 his from J. Broach; and A364A: MATa adel ade2 ural his7, lys2 tyri gall (KIL-o). Three new top] alleles created by gene disruptions are described in the text. The topl-6 allele contains the inserted LEU2 gene, the topl-7 allele contains deletion 1447-2295 as well as the LEU2 insertion, and the topl-8 allele contains deletion 580-1446 as well as the LEU2 insertion.Standard genetic methods and growth media were used (6). Transformation was by the spheroplast method (6). Two yeast genomic libraries were used, one was cloned in YCp5O, provided by M. Rose and G. Fink, and the other was cloned in YEp13 (7). DNA Topoisomerase I Assays. A method was developed by S. DiNardo of this laboratory whereby individual yeast colonies could be assayed directly for topoisomerase I activity without growth in liquid medium. A colony was scraped from a plate, resuspended in 30 ,ul of SED (1 M sorbitol/25 mM EDTA/50 mM dithiothreitol), and incubated at 370C for 30 min. Four microliters of zymolyase 60,000 (2.5 mg/ml) (Miles) was added. Cells were incubated at 30TC for 30 min. Spheroplasts were pelleted in a desktop centrifuge, resuspended in 12 ,ul of yeast lysis buffer (2), and kept on ice for 30 min. After a 10-min centrifugation, 1 gl of the supernatant could be assayed in a 10-,ul reaction mixture as described (1).Extracts for determining yeast topoisomerase I activity in E. coli were prepared by growing 20-m...

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Section: Agaagatgaagccaccattcacaagagaattattgatagagaaattgaaaaatatcagcgmentioning

confidence: 62%

Cloning, characterization, and sequence of the yeast DNA topoisomerase I gene.

Thrash¹,

Bankier²,

Barrell³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

174

105

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“…Although Baldwin et al (1983) have demonstrated that tyrosyl residues in human GH are phosphorylated by epidermal growth factor receptor in A431 cell membranes (Baldwin et al, 1983), it is unlikely that GH receptor-125I-GH complexes bound to the phosphotyrosyl binding antibody column via phosphotyrosyl residues in GH. 125I-GH by itself did not bind to the anti-phosphotyrosine antibody.…”

Section: Discussionmentioning

confidence: 99%

Growth hormone promoted tyrosyl phosphorylation of growth hormone receptors in murine 3T3-F442A fibroblasts and adipocytes

Foster¹,

Shafer²,

Rozsa³

et al. 1988

Biochemistry

115

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Because many growth factor receptors are ligand-activated tyrosine protein kinases, the possibility that growth hormone (GH), a hormone implicated in human growth, promotes tyrosyl phosphorylation of its receptor was investigated. 125I-Labeled human GH was covalently cross-linked to receptors in intact 3T3-F442A fibroblasts, a cell line which differentiates into adipocytes in response to GH. The cross-linked cells were solubilized and passed over a column of phosphotyrosyl binding antibody immobilized on protein A-Sepharose. Immunoadsorbed proteins were eluted with a hapten (p-nitrophenyl phosphate) and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The eluate from the antibody column contained an Mr 134,000 125I-GH-receptor complex. A similar result was obtained when the adipocyte form of 3T3-F442A cells was used in place of the fibroblast form. O-Phosphotyrosine prevented 125I-GH-receptor complexes from binding to the antibody column, whereas O-phosphoserine and O-phosphothreonine did not. In studies of GH-promoted phosphorylation in 3T3-F442A fibroblasts labeled metabolically with [32P]Pi, GH was shown to stimulate formation of a 32P-labeled protein which bound to immobilized phosphotyrosyl binding antibodies. The molecular weight of 114,000 obtained for this protein is similar to that expected for non-cross-linked GH receptor. The Mr 114,000 phosphorylated protein could be immunoprecipitated with anti-GH antibody, indicating that GH remained noncovalently bound to this protein during absorption to and elution from the immobilized phosphotyrosyl binding antibody. Phosphoamino acid analysis after both limited acid hydrolysis and extensive base hydrolysis of the Mr 114,000 phosphoprotein confirmed the presence of phosphotyrosyl residues.(ABSTRACT TRUNCATED AT 250 WORDS)

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confidence: 95%

“…As shown in Table 2 In addition, the introduction of the makl6-1 mutation improved the mating efficiency of cdcl6-1 strains 10-fold ( 25 acidic residues and no basic residues. Within the acidic region is a repeated sequence Asp-Asp-Gly-Glu-Val/GlyGlu-Tyr-Val, which is similar to sequences of known tyrosine phosphorylation sites (25). Serine residues at positions 212 and 241 and in the (Ser-Asp/Glu)7 alternating sequence from amino acids 251-264 are particularly good candidates for phosphorylation by casein kinases (26) that phosphorylate serine residues present in an acidic environment.…”

mentioning

confidence: 95%

Host function of MAK16: G1 arrest by a mak16 mutant of Saccharomyces cerevisiae.

Wickner

1988

Proc. Natl. Acad. Sci. U.S.A.

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The MAK16 gene was first defined as a gene whose mutation resulted in loss of M1 double-stranded RNA virus-like particles. The makl6-1 mutation also produces temperature-sensitive cell growth. We report here that makl6-I cells arrest at the nonpermissive temperature in GI phase, such that they are mating competent. We sequenced the MAK16 gene and found an open reading frame of 306 amino acids encoding a predicted protein ofMr 35,694. Two typical nuclear localization signal sequences were found. MAK16-LacZ fusion proteins that include one of these putative signals entered the nucleus, while unfused (3-galactosidase did not, as judged by subcellular fractionation experiments. In the C-terminal third of the MAK16 open reading frame is an acidic region in which 25 of 41 residues are either glutamate or aspartate. This region contains potential phosphorylation sites for "casein kinases," protein kinases specific for serine or threonine residues in an acidic environment.

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Phosphorylation of human growth hormone by the epidermal growth factor-stimulated tyrosine kinase.

Cited by 27 publications

References 32 publications

Cloning, characterization, and sequence of the yeast DNA topoisomerase I gene.

Cloning, characterization, and sequence of the yeast DNA topoisomerase I gene.

Growth hormone promoted tyrosyl phosphorylation of growth hormone receptors in murine 3T3-F442A fibroblasts and adipocytes

Host function of MAK16: G1 arrest by a mak16 mutant of Saccharomyces cerevisiae.

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