YARSI UNIVERSITY PROGRAM TO MEET THE DEMAND OF STUDENT’S ENGLISH FLUENCY WITH TOEIC AS THE ASSESSMENT TOOL

Abstract: A test is only a measurement tool of a learning process. The important part is the learning process itself; how the process can help learners acquire English as a foreign language that enables them to compete in the working environment. To measure the process, TOEIC with all its parts was meant to measure learners’ ability to communicate in English. Teachers should not be focusing on the test but more on the approaches that allow the students to have adequate and sophisticated listening, reading, and writing skills to exchange information and to negotiate meaning in real life. Many university level English teachers are trapped within the rules that students should achieve a 550 or 605 TOEIC score to graduate. Instead of helping the students to acquire the language as a communication tool, they tend to focus more on getting the students to master the test. This is what teachers should deal with, not only facilitate students to learn the language but at the same time help them to do the test well. Despite the challenge of facing students who lack motivation and have very basic English skills, Yarsi University Language Lab is setting up several programs and approaches that allow students to acquire the language and enable them to communicate in the target language which is eventually measured by an instrument called TOEIC. Keywords: Language Acquisition, direct and indirect test, discrete and intregativ

The increased retrotransposon mRNA levels in the <i>rpb1-CTD11</i> and <i>cdk8Δ</i> mutant were in part due to alterations to promoter activity.

<p>(A) Schematic of an average Ty1 promoter with binding sites for Gcn4 and Ste12/Tec1 labeled [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005608#pgen.1005608.ref048" target="_blank">48</a>]. TYA and TYB are retrotransposon genes that encode for the coat protein, and reverse transcriptase, protease, integrase, and RNase H respectively. YMLWTy1-2 and YJRWTy1-2 contain two Tec1 binding sites downstream of the ATG start codon, a feature observed on some Ty1 elements. Reporter assays for YMLWTy1-2 (B), YJRWTy1-2 (C), YDRWTy2-2 (D), and YLRWTy2-1 (E) in wild type, <i>rpb1-CTD11</i>, <i>cdk8Δ</i>, and <i>rpb1-CTD11 cdk8Δ</i>.</p

Genome-wide occupancy profiles of RNAPII suggested a role for the RNAPII-CTD in retrotransposon regulation.

<p>(A) Box plot showing differences in average MAT RNAPII (Rpb3) occupancy scores between the wild type and the <i>rpb1-CTD11</i> mutant at all, Ty1, or Ty2 retrotransposons. (B) Chromosome plots of relative RNAPII occupancy at representative retrotransposons. Increased RNAPII levels were observed in the <i>rpb1-CTD11</i> mutant compared to wild type. Labeled boxes indicate the retrotransposon. (C) Average gene profile of RNAPII occupancy at Ty1 retrotransposons showed increased levels along the length of the feature. Below, schematic of an average retrotransposon. Black triangles indicate the LTRs. (D) Average gene profile of RNAPII occupancy at Ty2 retrotransposons revealed increased levels towards the 3’ end of the feature.</p

The RNAPII-CTD Maintains Genome Integrity through Inhibition of Retrotransposon Gene Expression and Transposition

<div><p>RNA polymerase II (RNAPII) contains a unique C-terminal domain that is composed of heptapeptide repeats and which plays important regulatory roles during gene expression. RNAPII is responsible for the transcription of most protein-coding genes, a subset of non-coding genes, and retrotransposons. Retrotransposon transcription is the first step in their multiplication cycle, given that the RNA intermediate is required for the synthesis of cDNA, the material that is ultimately incorporated into a new genomic location. Retrotransposition can have grave consequences to genome integrity, as integration events can change the gene expression landscape or lead to alteration or loss of genetic information. Given that RNAPII transcribes retrotransposons, we sought to investigate if the RNAPII-CTD played a role in the regulation of retrotransposon gene expression. Importantly, we found that the RNAPII-CTD functioned to maintaining genome integrity through inhibition of retrotransposon gene expression, as reducing CTD length significantly increased expression and transposition rates of Ty1 elements. Mechanistically, the increased Ty1 mRNA levels in the <i>rpb1-CTD11</i> mutant were partly due to Cdk8-dependent alterations to the RNAPII-CTD phosphorylation status. In addition, Cdk8 alone contributed to Ty1 gene expression regulation by altering the occupancy of the gene-specific transcription factor Ste12. Loss of <i>STE12</i> and <i>TEC1</i> suppressed growth phenotypes of the RNAPII-CTD truncation mutant. Collectively, our results implicate Ste12 and Tec1 as general and important contributors to the Cdk8, RNAPII-CTD regulatory circuitry as it relates to the maintenance of genome integrity.</p></div

Truncation of the RNAPII-CTD resulted in altered association of a subset of transcription related factors at retrotransposons.

<p>Comparison of MAT average occupancy scores at Ty1 (A) or Ty2 (B) retrotransposons for the Mediator subunit, Cdk8, the mRNA capping enzyme, Cet1, the elongation factor, Elf1, the transcription elongation-associated chromatin mark, H3K36me3, and the general transcription factor, TFIIB, under wild type and <i>rpb1-CTD11</i> conditions.</p

Structural integrity of the RNAPII-CTD was important for normal retrotransposon mRNA levels and transposition rates.

<p>(A) Ty1 mRNA levels were significantly increased in the <i>rpb1-CTD11</i> mutant compared to wild type. (B) RT-qPCR analysis of Ty2 mRNA levels in the <i>rpb1-CTD11</i> mutant compared to wild type. RNAPII-CTD S₂ phosphorylation was important for maintaining normal Ty1 (C) and Ty2 mRNA levels (D). (E) Transposition rates for Ty1 were increased in the <i>rpb1-CTD11</i> mutant compared to wild type. Error bars represent 95% confidence intervals. Transposition rates and confidence intervals were calculated using the Fluctuation AnaLysis CalculatOR (FALCOR) web tool.</p

Increased levels of Ste12 at Ty1 and Ty2 promoters in the <i>cdk8Δ</i> mutant were normalized by truncation of the RNAPII-CTD.

<p>Box plot showing differences in average Ste12 (A) or Tec1 (B) occupancy scores between the wild type and the <i>rpb1-CTD11</i>, <i>cdk8Δ</i> and <i>rpb1-CTD11 cdk8Δ</i> mutant at Ty1 or Ty2 retrotransposons. Average gene profiles of Ste12 (C) or Tec1 (D) occupancy at Ty1 retrotransposons. Average gene profiles of Ste12 (E) or Tec1 (F) occupancy at Ty2 retrotransposons.</p

The increased Ty1 gene expression levels observed in the <i>rpb1-CTD11</i> mutant were dependent on <i>TEC1</i> or <i>STE12</i>.

<p>(A and B) Reporter assay for YMLWTy1-2 or YJRWTy1-2, with or without deletion of Tec1 binding sites. Tec1 binding sites were required for the increased promoter activity of Ty1 reporter constructs upon truncation of the RNAPII-CTD. (C and D) RT-qPCR analysis of wild type, <i>rpb1-CTD11</i>, <i>ste12Δ</i> and <i>rpb1-CTD11 ste12Δ</i> or wild type, <i>rpb1-CTD11</i>, <i>tec1Δ</i> and <i>rpb1-CTD11 tec1Δ</i> revealed that loss of <i>STE12</i> or <i>TEC1</i> in the <i>rpb1-CTD11</i> background led to Ty1 mRNA levels similar to those observed in the wild type.</p

A role for the RNAPII-CTD and Cdk8 in Ty1 gene expression regulation.

<p>Cdk8 normally functions to target Ste12 for degradation, thus loss of <i>CDK8</i> stabilized Ste12 at Ty1 promoters resulting in increased transcription initiation. Stability of Ste12 at Ty1 promoters was dependent on full length RNAPII-CTD, thus in the <i>rpb1-CTD11 cdk8Δ</i> double mutant Ste12 levels were normalized leading to wild type levels of transcription initiation. Truncation of the RNAPII-CTD alone resulted in increased Cdk8 recruitment to Ty1 elements and increased S₅ phosphorylation levels, a key mark for promoter clearance. The increased S₅ phosphorylation levels in the <i>rpb1-CTD11</i> mutant were direct or indirectly dependent on Cdk8, thus in the <i>rpb1-CTD11 cdk8Δ</i> double mutant S₅ phosphorylation levels were normalized resulting in decreased levels of promoter clearance.</p

Loss of <i>CDK8</i> normalized the elevated RNAPII and mRNA levels at Ty1 and Ty2 retrotransposons.

<p>(A) MAT average RNAPII (Rpb3) occupancy scores at retrotransposons revealed elevated levels at Ty1 and Ty2 elements in the single <i>cdk8Δ</i> mutant. In the <i>rpb1-CTD11</i> background, loss of <i>CDK8</i> resulted in normalized RNAPII levels at Ty1 and Ty2 retrotransposons. (B) Average gene profiles of RNAPII occupancy at Ty1 (top) or Ty2 (bottom) retrotransposons showed normalized RNAPII levels upon loss of <i>CDK8</i> in the <i>rpb1-CTD11</i> mutant. (C) Chromosome plots of RNAPII levels at representative retrotransposons. (D) Loss of <i>CDK8</i> normalized the elevated RNAPII levels at Ty1- and Ty2-derived LTRs observed in the <i>rpb1-CTD11</i> mutant. (E) RT-qPCR analysis of wild type, <i>rpb1-CTD11</i>, <i>cdk8Δ</i> and <i>rpb1-CTD11 cdk8Δ</i> revealed that loss of <i>CDK8</i> significantly normalized the elevated mRNA levels of Ty1 elements in the <i>rpb1-CTD11</i> background. (F) Ty2 mRNA levels were significantly elevated in the <i>cdk8Δ</i> mutant, an effect that was normalized when combined with an RNAPII-CTD truncation.</p