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L F Lau

Publications and source records attributed to L F Lau.

86 records · Page 5Linked to original sources
PubMed1989

A growth factor-responsive gene of murine BALB/c 3T3 cells encodes a protein homologous to human tissue factor.

Polypeptide growth factors rapidly induce the transcription of a set of genes that appear to mediate cell growth. We report that one of the genes induced in BALB/c mouse 3T3 cells encodes a transmembrane protein (mTF) homologous to human tissue factor, which is involved in the proteolytic activation of blood clotting. mTF mRNA is present in many murine tissues and cell lines. Our results raise the possibility that mTF may also play a role in cell growth.

Amino Acid Sequence↗
PubMed1988

A gene activated in mouse 3T3 cells by serum growth factors encodes a protein with "zinc finger" sequences.

We have recently identified by cDNA cloning a set of genes that are rapidly activated in mouse 3T3 cells by serum or purified growth factors. Here we report that the cDNA (clone 268) derived from one of these immediate early genes (zif/268) encodes a protein with three tandem "zinc finger" sequences typical of a class of eukaryotic transcription factors. The mRNA of zif/268 is present in many organs and tissues of the mouse and is especially abundant in the brain and thymus tissue. The 5' genomic flanking sequence of zif/268 has sequences related to binding sites for known regulatory proteins, including four sequences that resemble the core of the serum response elements (SREs) upstream of the c-fos and actin genes. The SRE-like sequences could be responsible for the coordinate activation of zif/268 and fos after serum stimulation of 3T3 cells.

Amino Acid Sequence↗
PubMed1988

A gene inducible by serum growth factors encodes a member of the steroid and thyroid hormone receptor superfamily.

We previously identified, by cDNA cloning, a set of genes that are expressed during the G0/G1 transition (cell cycle reentry) in mouse fibroblasts. These immediate early genes are transcriptionally activated within minutes of addition of serum or purified growth factors, and their mRNAs are superinduced in the presence of protein-synthesis inhibitors. We report here that one of these genes, represented by nur/77 cDNA (originally called 3CH77), encodes a member of the superfamily of ligand-binding transcription factors that includes the steroid and thyroid hormone receptors. The nur/77 cDNA sequence encodes a protein of 601 amino acids containing two regions of sequence similarity to members of this nuclear receptor superfamily, corresponding to their DNA-binding and ligand-binding domains. These results suggest that the growth factor-inducible immediate early gene nur/77 encodes a ligand-binding protein that regulates the genomic response to growth factors.

Amino Acid Sequence↗
PubMed1988

A gene activated by growth factors is related to the oncogene v-jun.

We have recently identified by cDNA cloning a set of genes that are rapidly activated in cultured mouse cells by protein growth factors. Here we report that the nucleotide sequence of a cDNA (clone 465) derived from one of these immediate early genes (hereafter called jun-B) encodes a protein homologous to that encoded by the avian sarcoma virus 17 oncogene v-jun. Homology between the jun-B and v-jun proteins is in two regions: one near the N terminus and the other at the C terminus. The latter sequence was shown by Vogt et al. [Vogt, P. K., Bos, T. J. & Doolittle, R. F. (1987) Proc. Natl. Acad. Sci. USA 84, 3316-3319] to have regions of sequence similarity to the DNA-binding domain of the yeast transcriptional regulatory protein GCN4 and to the oncogenic protein fos. Southern blots of human, mouse, and chicken DNA demonstrate that jun-B and c-jun are different genes and that there may be other vertebrate genes related to jun-B and c-jun. These findings suggest that there is a jun family of genes encoding related transcriptional regulatory proteins. The jun-B protein, and perhaps other members of the jun family, may play a role in regulating the genomic response to growth factors.

Amino Acid Sequence↗
PubMed1988

c-myc can induce expression of G0/G1 transition genes.

The human c-myc oncogene was linked to the heat shock-inducible Drosophila hsp70 promoter and used to stably transfect mouse BALB/c 3T3 cells. Heat shock of the transfectants at 42 degrees C followed by recovery at 37 degrees C resulted in the appearance of the human c-myc protein which was appropriately localized to the nuclear fraction. Two-dimensional analysis of the proteins of density-arrested cells which had been heat shock treated revealed the induction of eight protein species and the repression of five protein species. All of the induced and repressed proteins were nonabundant. cDNA clones corresponding to genes induced during the G0/G1 transition were used as probes to assay for c-myc inducibility of these genes. Two anonymous sequences previously identified as serum inducible (3CH77 and 3CH92) were induced when c-myc was expressed. In response to serum stimulation, 3CH77 and 3CH92 were expressed before c-myc mRNA levels increased. However, in response to specific induction of c-myc by heat shock of serum arrested cells, 3CH77 and 3CH92 mRNA levels increased after the rise in c-myc mRNA. Therefore, we hypothesize that abnormal expression of c-myc can induce genes involved in the proliferative response.

Animals↗
PubMed1987

Expression of a set of growth-related immediate early genes in BALB/c 3T3 cells: coordinate regulation with c-fos or c-myc.

We have previously identified by cDNA cloning 5 mRNAs that appear in resting BALB/c 3T3 cells soon after growth stimulation by serum or platelet-derived growth factor. Five additional mRNAs of this class are described in this report. The mRNAs reached peak levels between 40 and 120 min after serum addition and rapidly decayed thereafter. All 10 RNAs were superinduced in the presence of cycloheximide. Nuclear run-on experiments indicated that the increase in the mRNAs is the result of rapid transcriptional activation of their genes on stimulation by serum or platelet-derived growth factor. Superinducibility by cycloheximide is due to two effects: prolonged transcription and stabilization of mRNAs. This overall pattern of regulation is similar to that of the c-fos or c-myc protooncogenes reported previously. We hypothesize that these newly identified "immediate early" genes play a role in the proliferative response induced by growth factors.

Animals↗
PubMed1985

Identification of a set of genes expressed during the G0/G1 transition of cultured mouse cells.

To identify previously undetected genes that may be involved in the transition from a resting state (G0) to a proliferative state (G1) of mammalian cells, we set out to isolate cDNA clones derived from mRNAs that appear in serum-stimulated cells in the absence of protein synthesis. A lambda cDNA library was prepared using poly(A)+ RNA from BALB/c 3T3 cells that had been brought to quiescence and subsequently stimulated with serum in the presence of cycloheximide. Approximately 50 000 recombinant phage plaques were screened, and 357 clones were isolated that hybridized to probes derived from stimulated-cell RNA but not to probes from resting-cell RNA. Cross hybridization analysis showed that four RNA sequence families account for approximately 90% of these clones. One of the clones hybridized to an actin probe; none hybridized to any of 13 oncogene probes tested. Five different RNAs that appear to be previously uncharacterized have been further analyzed. These RNAs accumulate and decay rapidly following stimulation by serum or purified growth factors, or by a tumor promoter, and they are superinduced by serum in the presence of cycloheximide. Three of the RNAs could be enriched by hybridization to cDNAs and translated in vitro, yielding proteins of approximately 43, 40 and 35 kd, respectively.

Animals↗
PubMed1985

Rho-dependent transcription termination at lambda R1 requires upstream sequences.

In order to define the DNA sequence requirements for rho-dependent transcription termination, we have constructed and analyzed two classes of mutants affecting the cro-tR1 region of phage lambda: one class contains progressively larger deletions in the region upstream of the rho-dependent terminator tR1, and the other consists of small insertions or deletions generated by the linker scanning procedure. We show that rho-dependent termination at lambda tR1 in vitro requires contribution from sequences far upstream of the sites of termination. No single linker scanning mutant completely abolishes transcription termination, indicating that there exists no unique sequence that serves as an obligatory rho utilization site. Our results suggest that rho can interact with a large portion of the RNA transcript, including sequences that would be translated in vivo. Moreover, rho action appears to require a minimal transcript length, about 150-190 nucleotides for the tR1 termination sites. We also examine the effect of the auxiliary transcription factor NusA on transcription of these mutants, and we provide support for the proposal that specific effects of NusA on transcription termination require a sequence known as "boxA" (Olson, E.R., Flamm, E.L., and Friedman, D.I. (1982) Cell 31, 61-70).

Bacterial Proteins↗
PubMed1985

Phage lambda gene Q antiterminator recognizes RNA polymerase near the promoter and accelerates it through a pause site.

The positive regulator encoded by phage lambda gene Q is a transcription antiterminator that affects RNA polymerase initiating at the phage late gene promoter, but not at other promoters. We show that this nucleotide-sequence-specific interaction of Q protein and RNA polymerase can occur while the enzyme is pausing after 16 nucleotides of the late gene transcript have been made. Furthermore, Q protein chases RNA polymerase from this early pause site, so that it both recognizes the enzyme and changes its transcription properties at this site. We suggest that the ability of Q-modified RNA polymerase to escape this pause reflects the change that allows it to go through terminators. We also show that NusA protein is required for efficient Q protein activity in vitro.

Bacterial Proteins↗
PubMed1984

A potential stem-loop structure and the sequence CAAUCAA in the transcript are insufficient to signal rho-dependent transcription termination at lambda tR1.

It has been suggested that a sequence in the RNA transcript that can form a stem and loop structure, followed by the sequence CAAUCAA, is the signal for rho-dependent transcription termination. We tested this hypothesis by synthesizing a DNA duplex whose sequence corresponds to a region of the lambda tR1 terminator that contains these structural features. We cloned this synthetic DNA fragment under the control of the lacUV5 promoter, and showed that it does not cause rho-dependent termination in vitro. RNA polymerase pauses during in vitro transcription across the synthetic sequence, although less efficiently than at the corresponding sequence on the lambda template. No rho-mediated termination was detected even under conditions that prolonged transcriptional pausing at the synthetic site, indicating that the synthetic sequence is defective as a transcript release site. We suggest that unlike rho-independent terminators, rho-dependent terminators require sequences in addition to those immediately before the sites of termination.

Base Sequence↗
PubMed1983

RNA polymerase pausing and transcript release at the lambda tR1 terminator in vitro.

Transcription of the lambda rightward operon encounters multiple tandem p-dependent termination sites at a region known as tR1, yielding the mRNA for the lytic repressor cro. We show that in the absence of p factor, RNA polymerase pauses at each of three p-dependent termination sites (I, II, and III) at tR1 during in vitro transcription. We demonstrate that p factor itself does not affect transcriptional pausing at these sites. Relative occupancies of the pause sites during initial transcription have been measured. The lambda cin-1cnc-1 mutations, which abolish or vastly reduce termination at site II of tR1, nevertheless elicit transcriptional pausing at that site. The NusA protein (L factor) prolongs these transcriptional pauses to varying degrees. Transcriptional pausing at termination site II is dramatically enhanced by NusA in lambda wild type, but not in lambda cin-1cnc-1. NusA enhances pausing at site I but inhibits termination at that site, suggesting that it may also affect transcript release. We show that NusA does alter the kinetics of transcript release from isolated ternary complexes. We suggest that the ability of the NusA protein to increase or decrease termination efficiencies at various sites is attributed to its ability to modulate transcriptional pausing as well as transcript release.

Bacterial Proteins↗
PubMed1982

Transcription terminates at lambda tR1 in three clusters.

Transcription of the rightward operon of bacteriophage lambda terminates in the presence of rho factor at a region known as tR1, located downstream of the structural gene for the lytic repressor cro. We demonstrate by nuclease Sl mapping that transcription termination at lambda tR1 occurs in vitro over a stretch of 60 nucleotides. End points of transcripts are clustered in three distinct regions, which we refer to as termination sites I, II, and III. Termination at site I is inhibited by L factor, whereas termination at sites II and III is not affected by L factor. The sensitivities of these sites to rho factor are in the order III greater than II greater than I. The cin-lcnc-l mutations abolish termination at site II but not at sites I and III; this result may explain the failure of these mutations to alleviate the phage requirement for N function for growth. Although possible stem-and-loop structures in the RNA transcript immediately upstream of each of the three termination sites can be found, no consistent correlation exists between the strengths of these stem-and-loop structures and the termination efficiencies of their respective sites.

Bacteriophage lambda↗