Chapter 15, Problem 1VCQ

Figure 15.11 A scientist splices a eukaryotic promoter in front of a bacterial gene and inserts the gene in a bacterial chromosome. Would you expect the bacteria to transcribe the gene?

Summary Introduction

To analyze:

Whether the bacterial gene having eukaryotic promoter gets transcribed or not.

Introduction:

Eukaryotic promoters are different from prokaryotic promoters so the bacteria would not transcribe the gene.

Explanation of Solution

In case of transcription of genes, i.e. formation of RNA (mostly m-RNA) from DNA, promoters play an essential role. Promoters are the specific DNA sequences present near the starting point of transcription of a particular gene. Promoters help in the binding of RNA polymerase and transcription machinery to the specific point in the gene to initiate the process of transcription. They are mainly located upstream of a gene, towards 5’ end of the anti-sense strand. As these are specific recognition sequences, eukaryotic promoters are different from prokaryotic promoters.

Prokaryotic promoters have three essential elements:-10 elements,-35 element and UP (upstream promoter) element.

Eukaryotic promoters mainly contain a TATA box of six nucleotides (consensus sequence is 5’ TATAAA 3’) located around-30 (with reference to initiation site of transcription +1), while prokaryotic promoters have its counterpart-10 elements or Pribnow box of six nucleotides (consensus sequence is 5’ TATAAT 3’) located at-10. Both these have A-T rich elements. Prokaryotes also have one more promoter sequence of six nucleotides at-35 which is (5’ TTGACA 3’) called as-35 elements. Pribnow box or-10 element is essential in prokaryotes for the initiation of transcription and-35 element helps in increasing the transcription rate.UP element is A-T rich sequence located between-40 to-60.

Other conserved sequences in eukaryotic promoters are CAAT box (GGCCAATCT) around-80 and GC-rich boxes (GGCG), and octamer boxes (ATTTGCAT) located further upstream.Basically, eukaryotic promoters are much more complex and larger in structure than the prokaryotic promoters.Sigma factor (s factor) of prokaryotic RNA polymerase holoenzyme recognizes and binds to the promoter interaction site to initiate transcription.

So, a bacterial gene having eukaryotic promoter would not transcribe because eukaryotic promoter has different recognition sites/consensus sequences than the required prokaryotic recognition sites. RNA polymerase, transcription factors and other proteins of host bacterial chromosome would not be able to identify the eukaryotic promoter sequences and transcription of the gene would not occur.

Conclusion

So, the bacteria would not transcribe the gene with the eukaryotic promoter as they are different from the prokaryotic promoters.