DNA Directionality: 3' to 5' & Bacterial DNA Explained

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Discussion Overview

The discussion revolves around the directionality of DNA, specifically addressing the 3' to 5' and 5' to 3' orientations in the context of bacterial DNA, which is circular. Participants explore how directionality is determined in circular DNA and the implications for enzymatic processes such as replication and transcription.

Discussion Character

  • Exploratory
  • Technical explanation
  • Debate/contested

Main Points Raised

  • Some participants propose that directionality in DNA is based on the free end of the ribose, questioning how this applies to circular bacterial DNA where ends are joined.
  • Others argue that even in circular DNA, one strand runs 3'-5' while the complementary strand runs 5'-3', maintaining directionality.
  • A participant suggests that once the 3' end joins the 5' end, it becomes difficult to identify the starting points of the strands, leading to a discussion about the concept of direction versus ends.
  • Some participants raise questions about how polymerases can differentiate strands without cutting the DNA, indicating that this may depend on the context of replication or transcription.
  • It is noted that in replication, an enzyme may "nick" the DNA to create distinct ends, while in transcription, RNA polymerase binds to a promoter aligned in a specific direction.
  • One participant describes the perspective of directionality from the ribose ring structure, suggesting that the shape of the substrate can indicate direction without needing to cut the strand.

Areas of Agreement / Disagreement

Participants express differing views on how directionality is perceived in circular DNA and whether cutting is necessary to determine strand orientation. The discussion remains unresolved with multiple competing perspectives on the topic.

Contextual Notes

There are limitations in the discussion regarding assumptions about the definitions of sense and antisense strands, as well as the specific mechanisms by which polymerases recognize strand directionality.

at2341
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Hi,

To my understanding, the directionality of DNA (3' to 5') (5' to 3') is based on where the free(unattached) end of the ribose is.

With this idea, how can bacterial DNA (where the ends are joined together) have directionality?
 
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Cut it!
 
Circular DNA is still DNA. One strand will be 3'-5' and the other 5'-3'. Bases may only be added to the 3' end of any DNA strand, so that it is unidirectional.

To clarify: the 5' end is a phosphate grp and the 3' is a hydroxyl grp
 
Last edited:
I appreciate your replies,

Once the 3' end joins the 5' end to make a circle, you really can't tell where the 3' and 5' ends start. Is this correct? thanks
 
at2341 said:
Once the 3' end joins the 5' end to make a circle, you really can't tell where the 3' and 5' ends start. Is this correct? thanks


There is no start or end really, just direction. One strand will be in one direction, and the other complimentary strand in the other. As Matteo said...cut it! :)
 
1) Without cutting the DNA, there is no way to tell which strand is going in which direction? Is this correct?

2) If we can't tell which strand is which, how can the polymerase differentiate?

Thanks
 
I think what you may be asking about is which is the sense and anti sense strand.
If so then several steps are required for translation of a segment.
Unfolding, attachment of one or more promoters, and an available start codon.
AFAIK both sides of the DNA molecule can satisfy these conditions.
So the sense strand is the one that satisfies the requirements.
 
at2341 said:
1) Without cutting the DNA, there is no way to tell which strand is going in which direction? Is this correct?

2) If we can't tell which strand is which, how can the polymerase differentiate?

It depends on whether you are talking about replication (DNA polymerase) or transcription (RNA polymerase).

In replication, I believe there is an enzyme that "nicks" the DNA strand at a certain location (based on sequence) making a 3' and a 5' end. In transcription, the RNA polymerase binds to a promoter sequence which is aligned in a certain direction. Polymerase reads from 3' to 5' direction so that the new DNA or RNA it makes is building in the 5' to 3' direction.
 
thank you guys for replying.

BoomBoom, that's exactly what I wanted to know... how do the enzymes differentiate which way to go. thanks
 
  • #10
Consider a strand of DNA from the perspective of the middle of the ribose ring. The 3' direction is toward where the oxygen of a phosphate group hooks directly to the ring. The 5' direction is toward the extracyclic methylene which is bound to another phosphate. You don't need to cut the strand to see this directionality. Let's arbitrarily call one end of an enzyme's binding site the front; the enzyme would encounter a very different shape of substrate surface if it binds a strand of DNA with its front in the 3' direction as opposed to binding with its front in the 5' direction. No nick of the strand is needed, the shape of the ribose ring is enough to determine directions on the strand.
 
  • #11
JonMoulton said:
No nick of the strand is needed, the shape of the ribose ring is enough to determine directions on the strand.

Good point...I didn't think of that. Thanks Jon!
 

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