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C Montell

Publications and source records attributed to C Montell.

46 records · Page 3Linked to original sources

A rhodopsin gene expressed in photoreceptor cell R7 of the Drosophila eye: homologies with other signal-transducing molecules.

We have isolated an opsin gene from D. melanogaster that is expressed in the ultraviolet-sensitive photoreceptor cell R7 of the Drosophila compound eye. This opsin gene contains no introns and encodes a 383 amino acid polypeptide that is approximately 35% homologous to the blue absorbing ninaE and Rh2 opsins, which are expressed in photoreceptor cells R1-6 and R8, respectively. Amino acid homologies between these different opsins and other signal-transducing molecules suggest an important role for the conserved domains of rhodopsin in the transduction of extracellular signals.

Amino Acid Sequence↗

A second opsin gene expressed in the ultraviolet-sensitive R7 photoreceptor cells of Drosophila melanogaster.

We have identified a second Drosophila opsin gene, Rh4, which is expressed specifically in the ultraviolet-sensitive R7 photoreceptor cells. The two R7 opsins, Rh3 and Rh4, are expressed in nonoverlapping subsets of R7 cells. The Rh4 opsin shares 72% amino acid identity with Rh3, but is only about 35% homologous with the other 2 Drosophila opsins, ninaE and Rh2. Twenty-eight amino acids are conserved among all 4 invertebrate and 5 vertebrate opsin genes thus far sequenced, suggesting that these residues play an important role in rhodopsin function.

Amino Acid Sequence↗

Rescue of the Drosophila phototransduction mutation trp by germline transformation.

Phototransduction is the process by which light-stimulated photoreceptor cells of the visual system send electrical signals to the nervous system. Many of the steps that follow the initial event in phototransduction, absorption of light by rhodopsin, are ill-defined. The fruitfly, Drosophila melanogaster, provides a means to dissect phototransduction genetically. Mutations such as transient receptor potential (trp) affect intermediate steps in phototransduction. In order to facilitate molecular studies of phototransduction, the trp gene was isolated and its identity was confirmed by complementing the mutant trpCM allele of the trp gene by P-element mediated germline transformation of a 7.1-kilobase DNA fragment. Expression of the trp gene begins late in pupal development and appears to be limited to the eyes and ocelli.

Animals↗

Elimination of mRNA splicing by a point mutation outside the conserved GU at 5' splice sites.

Nearly all mRNA introns begin with the dinucleotide GU. Mutations in either of these virtually invariant bases have been found to inactivate the corresponding 5' splice site. Until now single base changes in neighboring bases have not been found to completely inactivate a 5' splice site. Here we show that a single A----U transversion in the third position of the adenovirus 2 E1A 13S mRNA intron does prevent RNA splicing at the corresponding 5' splice site.

Adenoviruses, Human↗

Complete transformation by adenovirus 2 requires both E1A proteins.

Rodent cells transformed by adenovirus 2 (Ad2) express two highly related viral proteins of 289 and 243 amino acids encoded in early region 1A (E1A). Transformation studies were performed with adenovirus mutants that express only one or the other E1A protein. We found that the 289 amino acid protein, which has transcription inducing activity, and the 243 amino acid protein, which has little if any of this activity, were both required to produce the fully transformed phenotype. Expression of either E1A protein induced a partially transformed phenotype. The 243 amino acid protein was particularly important for anchorage independent growth. As found in previous studies with several other E1A mutants, the process of transformation by the mutant that expresses the 243 amino acid protein only was cold-sensitive. While the 289 amino acid protein is the only E1A protein required for efficient viral replication under standard cell culture conditions, the 243 amino acid protein in addition to the 289 amino acid protein was found to be required for efficient viral replication in growth-arrested human cells.

Adenoviruses, Human↗

Control of adenovirus E1B mRNA synthesis by a shift in the activities of RNA splice sites.

The primary transcript from adenovirus 2 early region 1B (E1B) is processed by differential RNA splicing into two overlapping mRNAs, 13S and 22S. The 22S mRNA is the major E1B mRNA during the early phase of infection, whereas the 13S mRNA predominates during the late phase. In previous work, it has been shown that this shift in proportions of the E1B mRNAs is influenced by increased cytoplasmic stability of the 13S mRNA at late times in infection. Two observations presented here demonstrate that the increase in proportion of the 13S mRNA at late times is also regulated by a change in the specificity of RNA splicing. First, the relative concentrations of the 13S to 22S nuclear RNAs were not constant throughout infection but increased at late times. Secondly, studies with the mutant, adenovirus 2 pm2250 , provided evidence that there was an increased propensity to utilize a 5' splice in the region of the 13S 5' splice site at late times in infection. Adenovirus 2 pm2250 has a G----C transversion in the first base of E1B 13S mRNA intron preventing splicing of the 13S mRNA but not of the 22S mRNA. During the early phase of a pm2250 infection, the E1B primary transcripts were processed into the 22S mRNA only. However, during the late phase, when the 13S mRNA normally predominates, E1B primary transcripts were also processed by RNA splicing at two formerly unused or cryptic 5' splice sites. Both cryptic splice sites were located much closer to the disrupted 13S 5' splice site than to the 22S 5' splice site. Thus, the temporal increase in proportion of the 13S mRNA to the 22S mRNA is regulated by two processes, an increase in cytoplasmic stability of the 13S mRNA and an increased propensity to utilize the 13S 5' splice site during the late phase of infection. Adenovirus 2 pm2250 was not defective for productive infection of HeLa cells or for transformation of rat cells.

Adenoviruses, Human↗

Resolving the functions of overlapping viral genes by site-specific mutagenesis at a mRNA splice site.

Early region IA of human adenoviruses encodes a function required for normal induction of early viral genes and virus-induced cell transformation. The region is expressed at early times as two overlapping spliced mRNAs, 12S and 13S, which encode closely related proteins. To distinguish between the functions of these proteins, a single T leads to G transversion was constructed which prevents splicing of the 12S mRNA. This transversion, in the second base of the 12S mRNA intron, does not alter the protein encoded by the 13S mRNA due to degeneracy in the genetic code. Studies with this mutant demonstrated that only the 13S mRNA encodes the regulatory protein required for normal early gene expression.

Adenoviruses, Human↗

Enhanced expression of adenovirus transforming proteins.

Proteins encoded in regions EIA and EIB of human adenoviruses cause transformation of rodent cells. One protein from EIA also stimulates transcription of other early regions at early times in a productive infection. In the past, direct analysis of these proteins synthesized in vivo has been difficult because of the low levels produced in both transformed cells and productively infected cells. We present a simple method which leads to expression of EIA and EIB mRNAs and proteins at 30-fold greater levels than those observed during the early phase of a standard productive infection. Under these conditions, these proteins are among the most prominent translation products of infected cells. This allowed direct visualization of EIA and EIB proteins on two-dimensional gels of pulse-labeled total cell protein. Experiments with EIA and EIB mutants confirm that the identified proteins are indeed encoded in these regions. Two EIA proteins are observed, one translated from each of the major early EIA mRNAs. Both of these EIA proteins are phosphorylated.

Adenoviruses, Human↗

Molecular genetics of Drosophila vision.

The fruitfly, Drosophila melanogaster, is an excellent organism for dissecting the components of vision genetically. Many mutations have been generated that affect a diversity of processes important in vision. Through a combined application of molecular and genetic approaches many of the genes important in Drosophila vision are now being identified.

Animals↗

Inhibition of RNA cleavage but not polyadenylation by a point mutation in mRNA 3' consensus sequence AAUAAA.

A single U leads to G transversion in the 3' consensus sequence AAUAAA of the adenovirus early region 1A gene was constructed and the effect of this mutation on processing of the 3' end of the nuclear early region 1A RNAs was analysed. The results demonstrate that the intact AAUAAA is not required for RNA polyadenylation but is required for the cleavage step preceding polyadenylation to occur efficiently.

Adenoviruses, Human↗