Cloning and sequence analysis of cDNA for rat spleen thymosin beta 4.

Wodnar-Filipowicz, Aleksandra; Gubler, Ueli; Furuichi, Yasuhiro; Richardson, Michael; Nowoswiat, Eugene F.; Poonian, Mohindar S.; Horecker, B.L.

doi:10.1073/pnas.81.8.2295

Cited by 33 publications

(21 citation statements)

References 17 publications

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“…By this approach, 9 of the 10 clones showed no significant homologies to known sequences. However, clone 21b was found to be highly homologous to thymosin P34, a peptide with a wide tissue distribution (23,69). Clone 21b includes the entire coding region of thymosin 134, has an additional 21 bp at the 5' end, and extends to nucleotide 419 of the 523-bp published cDNA sequence (23) (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Thymosin 4 mRNA is induced by alpha interferon in a human neuroblastoma cell line (21) and a lymphoblastoid cell line (49). Wodnar-Filipowicz et al (69) have hypothesized that thymosin N may be a component of the cytoskeleton. While such a role for thymosin 4 iS intriguing given the extensive morphological changes involved in PC12 neuronal differentiation and the induction of this mRNA by NGF, the precise physiologic role of thymosin 4 in NGF-responsive as well as in other cell types remains to be determined.…”

Section: Discussionmentioning

confidence: 99%

“…This led to the proposal that thymosin 4 is a secreted thymic hormone (46). However, thymosin 4 mRNA does not encode a signal peptide (23,69). Thymosin N protein (32,33,70) and mRNA (Fig.…”

Section: Discussionmentioning

confidence: 99%

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Identification and characterization of mRNAs regulated by nerve growth factor in PC12 cells.

Leonard¹,

Ziff²,

Greene³

1987

Mol. Cell. Biol.

281

137

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Differential screening of cDNA libraries was used to detect and prepare probes for mRNAs that are regulated in PC12 rat pheochromocytoma cells by long-term (2-week) treatment with nerve growth factor (NGF). In response to NGF, PC12 cells change from a chromaffin cell-like to a sympathetic-neuron-like phenotype. Thus, one aim of this study was to identify NGF-regulated mRNAs that may be associated with the attainment of neuronal properties. Eight NGF-regulated mRNAs are described. Five of these increase 3-to 10-fold and three decrease 2-to 10-fold after long-term NGF exposure. Each mRNA was characterized with respect to the time course of the NGF response, regulation by agents other than NGF, and rat tissue distribution. Partial sequences of the cDNAs were used to search for homologies to known sequences. Homology analysis revealed that one mRNA (increased 10-fold) encodes the peptide thymosin 4 and a second mRNA (decreased 2-fold) encodes tyrosine hydroxylase. Another of the increased mRNAs was very abundant in sympathetic ganglia, barely detectable in brain and adrenals, and undetectable in all other tissues surveyed. One of the decreased mRNAs, by contrast, was very abundant in the adrenals and nearly absent in the sympathetic ganglia. With the exception of fibroblast growth factor, which is the only other agent known to mimic the differentiating effects of NGF on PC12 cells, none of the treatments tested (epidermal growth factor, insulin, dibutyryl cyclic AMP, dexamethasone, phorbol ester, and depolarization) reproduced the regulation observed with NGF. These and additional findings suggest that the NGF-regulated mRNAs may play roles in the establishment of the neuronal phenotype and that the probes described here will be useful to study the mechanism of action of NGF and the development and differentiation of neurons.Nerve growth factor (NGF) plays a major role in the growth, differentiation, and survival of sympathetic and sensory neurons (22,41). Recent studies have demonstrated the presence of NGF and its receptor in the central nervous system and have shown that certain neurons of the forebrain respond to NGF (35,51). One in vitro model system used to study the effects and mechanism of action of NGF is the rat PC12 pheochromocytoma cell line (27,28). In the presence of NGF, PC12 cells differentiate from replicating, chromaffinlike cells to nonreplicating, sympathetic-neuron-like cells. One of the more dramatic aspects of PC12 cell differentiation is the attainment of a neuronal phenotype by the extension of long, branching neurites. Within 1 week of NGF treatment, approximately 90% of the cells have neurites. NGF-treated PC12 cells also develop the ability to generate fast, Na+-dependent action potentials (13) and undergo alterations in the levels of a numDer of specific proteins, many of which change in an RNA synthesis-dependent manner (see, for example, reference 28 for review). Burstein and Greene (6) demonstrated that both RNA synthesis-dependent and -independent events are necessary for neurite o...

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Identification and characterization of mRNAs regulated by nerve growth factor in PC12 cells.

Leonard¹,

Ziff²,

Greene³

1987

Mol. Cell. Biol.

281

137

View full text Add to dashboard Cite

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“…Endogenous Tb4 is known to be enzymatically acetylated in the cytosol. 34 The results showed that this PTM occurs in 293F cells.…”

Section: Membrane Permeabilizationmentioning

confidence: 92%

CHAPTER 4. In‐Cell NMR Spectroscopy to Study Protein–Drug Interactions

Washington¹,

Burz²,

Shekhtman³

2013

New Developments in NMR

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“…1 Three of the 16 purified cDNA sequences appeared identical to previously characterized genes that were not known to be expressed in pre-B-cells. The most highly represented sequence (designated PB37) among the original 97 phage clones corresponded to the previously defined gene for thymosin (34), a small secreted protein of unknown function. The PB17 sequence, which was represented once, corresponded to the gene for type II carbonic anhydrase (35).…”

Section: Resultsmentioning

confidence: 99%

Isolation of coordinately regulated genes that are expressed in discrete stages of B-cell development.

Yancopouloš

Oltz

Rathbun

et al. 1990

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

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We have utilized subtractive hybridization to isolate 16 distinct cDNA sequences representing genes expressed in pre-B-cell lines but not myeloma cell or fibroblast lines. These sequences represent RNA transcripts that vary in abundance in pre-B-cell lines from 0.001% to 0.05%. Five of these sequences were not related to any known genes. One was related to but distinct from known myosin regulatory light chain genes and another encoded a protein with lectin domains. Three represented previously identified genes encoding carbonic anhydrase type II, thymosin, and CD2; these genes were not previously known to be specifically expressed in early stages of B-cell development. Other isolated genes corresponded to pre-B-cell-specific or pre-B-cell/B cell-specific genes recently described by others. The isolated cDNA sequences may be divided into two general categories-those representing genes expressed only in the pre-B-cell stage ofB-cell development and those expressed in both the pre-B-cell and B-cell stages. The in vivo expression patterns of the identified genes suggest that some function specifically in lymphocytes while others may have roles in additional lineages.Differentiation of B-lineage cells occurs in three general stages-the precursor B-cell (pre-B-cell) stage, the Blymphocyte stage, and the plasma-cell stage (1). In mammals, pre-B-cell development occurs in the liver of the fetus and shifts to bone marrow in adults. Pre-B-cells first assemble heavy (H) chain variable (V) region genes from component variable (VH) diversity (D), and joining (JH) gene segments and subsequently assemble light (L) chain V region genes from VL and JL segments (2). Expression of H and L chains by pre-B-cells leads to the generation of B lymphocytes, which express complete immunoglobulin molecules on their surface (3). B lymphocytes migrate to peripheral lymphoid organs, such as the spleen and lymph nodes (1). MATERIALS AND METHODSCell Lines. The various cell lines and their characteristics have been described (11,12,(22)(23)(24)(25)(26)(27).cDNA Preparation. Radiolabeled cDNA of relatively low specific activity was made from 6 ,Ag ofpoly(A)+ RNA pooled from three pre-B-cell lines (38B9, 22D6, and 300-19P) as described (28).Preparative and Analytical Hybridizations. Details of the subtractive hybridizations, S1 nuclease assays, and fractionation of double-and single-stranded nucleic acids by fractionation on hydroxyapatite were essentially as described (28). The cDNA from the above reaction was added to 50 pg ofpoly(A)+ RNA from the Ltk-fibroblast cell and hybridized to an Rot of about 5000. After hybridization, single-stranded cDNA (about 5-10%o of the input cDNA) was isolated by hydroxyapatite chromatography and was hybridized back to 50 ug ofpre-B-cell poly(A)+ RNA (to an Rot of5000) to render it double-stranded; the fraction of this cDNA that hybridized back to autologous RNA was estimated to be about 60% by S1 nuclease analysis. The cDNA*RNA hybrids were used

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Cloning and sequence analysis of cDNA for rat spleen thymosin beta 4.

Cited by 33 publications

References 17 publications

Identification and characterization of mRNAs regulated by nerve growth factor in PC12 cells.

Identification and characterization of mRNAs regulated by nerve growth factor in PC12 cells.

CHAPTER 4. In‐Cell NMR Spectroscopy to Study Protein–Drug Interactions

Isolation of coordinately regulated genes that are expressed in discrete stages of B-cell development.

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